Sci Eng Ethics DOI 10.1007/s11948-015-9628-2 ORIGINAL PAPER

The Acid Test for Biological Science: STAP Cells, Trust, and Replication Cheryl Lancaster

Received: 31 October 2014 / Accepted: 26 January 2015  Springer Science+Business Media Dordrecht 2015

Abstract In January 2014, a letter and original research article were published in Nature describing a process whereby somatic mouse cells could be converted into stem cells by subjecting them to stress. These ‘‘stimulus-triggered acquisition of pluripotency’’ (STAP) cells were shown to be capable of contributing to all cell types of a developing embryo, and extra-embryonic tissues. The lead author of the publications, Haruko Obokata, became an overnight celebrity in Japan, where she was dubbed the new face of Japanese science. However, in the weeks that followed publication of the research, issues arose. Other laboratories and researchers (including authors on the original papers) found that they were unable to replicate Obokata et al.’s work. Closer scrutiny of the papers by the scientific community also suggested that there was manipulation of images that had been published, and Obokata was accused of misconduct. Those who should have been supervising her work (also her co-authors on the publications) were also heavily criticised. The STAP cell saga of 2014 is used as an example to highlight the importance of trust and replication in twenty-first century biological science. The role of trust in the scientific community is highlighted, and the effects on interactions between science and the public examined. Similarly, this essay aims to highlight the importance of replication, and how this is understood by researchers, the media, and the public. The expected behaviour of scientists in the twenty-first century is now more closely scrutinised. Keywords

STAP cells
Replication
Trust
Misconduct
Haruko Obokata

C. Lancaster (&) Department of Philosophy, Durham University, 50 Old Elvet, Durham DH1 3HN, UK e-mail: [email protected]

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Introduction Misconduct in science is certainly not a new phenomenon. For example, in the 1970s, William Summerlin blackened patches of fur on white mice where he was supposed to have carried out skin transplants from black to white mice. In 2003, Steven A Leadon at the University of North Carolina fabricated data regarding his research into DNA repair. The virologist Naoki Mori from the University of Ryukyus in Japan was also accused of manipulating data and images in several publications. A blooming field in biological sciences research, stem cell biology, has also had its share of misconduct. For instance, in 2009, Jizhen Lin at the University of Minnesota was accused of manipulating figures in a 2008 paper describing how stem cells from the inner ear could become neurons. In late January 2014, two publications appeared in Nature describing the creation of stem cells using relatively simple techniques. These so-called stimulus-triggered acquisition of pluripotency (STAP) cells were created from murine cells, and suggested that any cell, under appropriate conditions, could be ‘re-programmed’ to become a cell capable of contributing to all cells of an adult mouse. This was initially considered to be a very important advancement: the relatively easy technique the article described suggested that any mammalian adult cell could be converted into a stem cell, with the potential to re-differentiate into any other cell type. Reactions from the scientific community were positive for a few days, however a week following publication, questions began to be raised. In particular, these focused on the accuracy of some of the figures published, and the inability of others to replicate the results using the methods provided. As concerns arose regarding the possibility of fraud, the lead institute (where significant amounts of experimental work for these papers had been carried out), RIKEN, Japan, began an investigation. In a flurry of Japanese and worldwide press interest, RIKEN announced that the lead author of both publications, Haruko Obokata, was guilty of misconduct; that the STAP cells she claimed to have produced had not been created at all. The question arises, why are there well-known cases of fraud in stem cell biology? Does it occur with the same frequency in other research areas, yet goes undetected? Or is it not interesting enough to be reported by mainstream media? The episode has given the general public an in-depth insight into how science operates. The STAP cell case suggests that there are issues in biological science research including slovenly peer review, poor supervision and oversight, and inadequate reporting in the media. This essay suggests that this can affect the public trust in science, as well as hinder scientific progress.

Fraud in Biological Sciences: A Retrospective Published Fraud In 2012, Fang, Steen, and Casadevall published a paper in PNAS concerning the number of papers retracted for fraud, plagiarism, or duplicated publication

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(misconduct) in the biomedical and life sciences (Fang et al. 2012). Between 1973 and 2012, there were 2047 articles listed on the PubMed database1 as ‘retracted’. Fang, Steen, and Casadevall report that 43.3 % of these papers were retracted due to fraud or suspected fraud, with an almost tenfold increase in the number of retractions because of fraud since 1975. (This recent increase was not solely due to the increased number of research publications.) With regards to stem cell fraud, and its frequency, there has (as yet) been no indepth study such as that carried out by Fang, Steen, and Casadevall. This makes it difficult to put a number or percentage on the number of retractions (for fraud) for stem cell biology in comparison to the wider biological sciences field. Using a very basic version of the methods in Fang, Steen, and Casadevall, PubMed was searched for all articles containing the term ‘‘stem cell’’ between 1973 (when Fang, Steen, and Casadevall report the first retraction) and May 2012 (when Fang, Steen, and Casadevall carried out their search); this resulted in 102,089 hits. When an additional search for retractions was added, this resulted in 62 hits; 0.06 % of papers referring to stem cells were therefore retracted between 1973 and mid-2012. A brief check of retraction notices sheds light on the reasons for these retractions. Considering Fang, Steen, and Casadevall’s definition of fraud, 25 of 59 retractions (with reasons given) were for fraud: 42.37 %, a percentage equivalent to that claimed for all biomedical and life sciences. In the academic press then, stem cell fraud does not appear to occur with more frequency than in general bioscience. Does the reporting of fraud in stem cell biology in the popular press occur with more frequency? This is a much more difficult to assess. Searches of databases can not demonstrate with the same accuracy the number of popular press articles, blog posts, tweets etc. that refer to any scientific fraud, nor will it give an indication of the number of people who have read them. The number of reports regarding single cases of fraud however may be indicative of public interest: newspapers would not continue to print news articles about topics their readership was not interested in. A brief search of the archives of two newspapers, The Guardian, a UK newspaper, and The New York Times, a more internationally-recognised newspaper, reveals results along these lines. Searches for ‘Naoki Mori’ and ‘Steven Leadon’ reveal 5 articles between the two newspapers. Searches for ‘Hwang Woo Suk’ reveals 170 in The Guardian and 391 in The New York Times. A search for ‘Haruko Obokata’, shows 23 articles in The Guardian, and 30 in The New York Times. These basic searches suggest that the popular press are more likely to report and continue reporting on stem cell misconduct rather than misconduct in other areas of biological science. Public trust and hinderance of scientific progress In 2000 a House of Lords Select Committee in the UK considered the interaction between science and society. Whilst concluding that public interest in science in the UK is high, there is negativity associated with science whose purpose is not obviously beneficial, and mistrust of governmental or industrial associated science. 1

PubMed is a database of primarily biomedical sciences research articles. It is maintained by the US National Library of Medicine at http://www.ncbi.nlm.nih.gov/pubmed/.

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There is a perception of a governmental and industrial secrecy culture, which invites suspicion. Such negative associations are expressed as lack of trust; suppressing uncertainty is expected to erode public trust. With this in mind, the Select Committee suggested that scientists are becoming more responsible for the public understanding of science. Nevertheless, the report states, the present ‘crisis of trust’ has ‘political reality’. Why does public trust in science matter? Firstly, the future of any society depends significantly on young people; without young people interested in pursuing careers in science, the Select Committee were concerned that the welfare and wealth of society could be critically affected. The applications of current science also feed into society, often via policy and industry, which are also vulnerable to public trust. Technologies and research that do not have public confidence (such as genetically modified crops or therapeutic cloning), regardless of the reasons for such mistrust, can inhibit funding and policy to allow further research into controversial areas. Public health may also suffer where there is mistrust and misunderstanding—for instance, inappropriate use of antibiotics has led to bacterial resistance. Politics, industry, and science then can all be affected by public opinion and trust.

Case Study: Reports on ‘‘Stimulus-Triggered Acquisition of Pluripotency’’ The STAP Cell Publications On 29 January 2014, a letter (Obokata et al. 2014b) and an original research article (Obokata et al. 2014a) were published in the highly-regarded journal Nature. The article, ‘‘Stimulus-triggered fate conversion of somatic cells into pluripotency’’, described methods by which normal cells of the mouse could be turned into cells with an important property: the ability to become any other cell of the body. Up until this point, the only cells known to be able to achieve this were embryonic stem cells, or some induced pluripotent stem cells (iPS cells). Inducing pluripotency is a rather complex procedure,2 whereas the Obokata et al. Nature article suggested that subjecting cells to stress (such as exposure to weak acid3) could cause them to regain their ability to become a variety of cell types; a much simpler method than that used to create iPS cells. The group, led by researchers at the RIKEN Center for Developmental Biology in Japan, argued that ‘‘stimulus-triggered acquisition of pluripotency’’ (STAP) could indicate that fate determination of mammalian cells 2

The initial procedure of creating induced pluripotent stem cells was carried out by Shinya Yamanaka’s laboratory at Kyoto University in 2006 (see Takahashi and Yamanaka 2006). The procedure required the introduction of four factors to adult fibroblast cells, which were cultured in conditions usually used to grow embryonic stem cells. These cells were eventually shown to be capable of contributing to all cells of the murine embryo. The protocol for creating mouse induced pluripotent cells is around 1–2 weeks, and is relatively inefficient.

3 Several techniques are described in Obokata et al. (2014a) for stressing the cells. These include: treating cells in a low-pH (pH 5.7) solution for 25 min at 37 C; titrating cells through narrow pipettes (50 um) for 20 min; heating the cells to 42 C for 20 min; nutrient deprivation for three weeks; growing in high calcium (2 mM) media for 2 days; and creating pores in the cell membrane by treating cells with streptolysine O for 2 h. It was the acid treatment that was the most successful method.

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can be influenced by environmental cues.4 The letter submitted and printed in the same edition of Nature by a similar group (also led by researchers at RIKEN), went a step further, suggesting the STAP cells were actually capable of contributing to bodily tissues and so-called ‘extra-embryonic’ tissues in mice.5 These experiments were carried out using mouse cells, and Obokata et al. claimed that mice had been created using mouse STAP cells. In Japan, the lead author of both the article and letter, Haruko Obokata, became an overnight celebrity. Obokata, a 30-year old woman, stood out against the stereotypical image of the ageing male scientist; her breakthrough saw her hailed as the new face of Japanese science. The Japanese media singled Obokata out, and reported on her consistently in the days following the publication of her work. Consequently, the clothes she wore at the press conference announcing publication of the STAP cell papers, sold out across the country. Both papers were received by Nature in the middle of March 2013, and accepted in December 2013. During this time, presumably the usual peer review process occurred, whereby Nature editors sent the papers to various experts in the field and asked for comments, particularly regarding the papers’ suitability for publication. Apparently, after 9 months, the reviewers and editors agreed that the papers were acceptable for publication in Nature. It is now known that in addition to this, similar articles had been submitted to two other journals, Cell, and Science. Both had rejected the paper; initially, Nature had also rejected a manuscript. Reviewers of the Cell submission (including Hans Scho¨ler, a stem cell researcher at the Max Planck Institute for Molecular Biomedicine in Germany) were concerned about the lack of evidence presented for such extraordinary claims. Science reviewers observed issues with figures (Normile and Vogel 2014). Reactions from the Scientific Community The initial reaction from the stem cell community was that this was a useful breakthrough. It provided further evidence that environmental cues could affect differentiation (a technique that has been used successfully in the laboratory for several years), and in addition presented a new ‘quick and easy’ method of creating cells that could turn into almost any other. The work was considered exciting, if surprising, although most agreed that more work was needed, particularly as the authors could not yet explain the mechanism by which stress could cause cells to revert back to a stem cell-like state. Further opinion seemed confused as to why the cells in the article did not appear to have a stem cell morphology; it was generally agreed though that morphology was not at this time a key issue if the cells were expressing the appropriate proteins. 4

It is currently understood that the stem cell environment is important for maintaining cells as stem cells, and inducing differentiation. For example, mechanical (such as cell adherence to other cells or extracellular matrix, for instance) and chemical (exposure to extracellular ‘factors’) cues can affect cell division and differentiation. This function works in tandem with expression of certain genes.

5

These are tissues such as the placenta, that are required by the foetus for development but do not form part of the body. These cells are referred to as ‘totipotent’.

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An interview with Charles A. Vacanti, an author on the STAP publications, and a mentor of Obokata at Harvard Medical School, appeared online on 2 February 2014. Vacanti was able to answer some questions other researchers had about the process of creating STAP cells. Vacanti claimed that he still could not explain the mechanism of STAP, however believed that demethylation of some genes may have been an important factor.6 Answering queries regarding cell stress and environmental conditions, Vacanti highlighted that although some regions of the body had, for example, low pH environments, in other regions, cells are not equipped to survive in such environments. Hardier mature cells then revert to a stem cell-like state, allowing regeneration and repair of the region. Furthermore, Vacanti claimed that human STAP cells had been created, however these experiments were at an early stage, and work was ongoing (Knoepfler 2014). On 5 February 2014, Vacanti sent New Scientist an image of what were allegedly human STAP cells; human fibroblasts were stressed (Vacanti would not reveal how at this point), and resulted in cells similar to the mouse STAP cells reported in Obokata et al. (2014a), Thomson (2014). A week after publication however, at the beginning of February 2014, concerns were starting to be raised. Firstly, the method seemed a little too simple—why had something like this not been achieved before if undifferentiated cells could be created from simple procedures such as treating cells with acid? Secondly, despite the apparent simplicity and speed of the technique, no other researcher had yet been able to replicate the results. But this was just a few days after the work was published—perhaps skill was slightly lacking or the nuances of the methods were not described in the publications. Whatever the reason, lack of replication after only a week is not a particularly strong reason to entirely dismiss work that has been peer reviewed and published in Nature. On 4 February 2014, the first comment about figure manipulation was suggested on the website PubPeer.7 The first comment referred to Figure 1i in Obokata et al. (2014a), which depicted a PCR analysis. There was concern that a section of this image had been spliced in.8 General thoughts were that this was not an appropriate way of presenting data, and that it suggested sloppiness at the least. The following day, further comments were made on PubPeer, concerned about the appearance of some of the immunofluorescence images in the article. In particular, there was concern that autofluorescence was occurring (which is usual in dead or dying cells), and that the images had been

6

‘Demethylation’ is a process that changes the molecular structure of DNA. This occurs normally, and can occur across the entire genome (for example, in paternal DNA immediately following fertilisation) or in a specific region of DNA. Demethylation (and its opposite counterpart, methylation) is part of a cell’s mechanism to influence which genes are ‘turned on’ and which are ‘turned off’. (For a detailed review, see Chen and Riggs 2011.)

7

PubPeer is a website that involves an online community of scientists, giving them a platform to discuss scientific articles. Authors are encouraged to join discussions of their research papers.

8

In this instance, Obokata had carried out PCR and had run gels to check the results of the PCR. Instead of showing that these gels were run at different times (by including two separate images), Obokata had edited the gel images to appear as though the results were produced at the same time (i.e. there was one gel showing the results of a PCR that occurred on one occasion). This was considered to be misleading.

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misinterpreted. There was further suspicion that the same image had also been used to show apparently different placentas in the published letter.9 A week following publication, calls were made to ‘crowdsource’ replication attempts. A fortnight later no-one could report that they had successfully replicated the results, despite several attempts from different laboratories. This suggested that the technique, at best, might not be as simple as originally thought. Contrary to Nature policy, the authors had not yet deposited any of their experimental data for interested people to view.10 Concerns also began to be raised about Obokata’s previously published work. Three weeks post-publication, a news piece in Nature, apparently unable to ignore the controversy any longer, admitted asking ten laboratories to assess the methods presented in the STAP cell papers; none were able to reproduce the results published. In addition, one of the original authors (Teruhiko Wakayama) claimed that he was unable to get the same results in a different laboratory. Investigation With Nature apparently admitting that there may be an issue with the publications, the lead institution RIKEN began an investigation. In response to this, Obokata, Yoshiki Sasai, and Hitoshi Niwa published ‘tips’ for successful replication of the STAP cell technique online, at the Nature Protocol Exchange on 3 March. Meanwhile, Waseda University in Japan, where Obokata obtained her PhD, also began investigating her work. Studies of Obokata’s thesis revealed that claims of results of different experiments were actually the same image. There were also significant plagiarism issues arising from Obokata’s thesis, including significant sections of her introduction and references; Obokata offered to withdraw her thesis on 15 March. The Japanese press were particularly concerned with the claims that Obokata had manipulated some of the figures presented in her work (that no other laboratory had yet reproduced the results was not an issue). Wakayama, a senior author on the publications, began to call for the retraction of the original research article. In an email to The Wall Street Journal, Wakayama stated that ‘‘there is no more credibility when there are such mistakes’’ (Martin and Naik 2014). Eventually, on 14 March, RIKEN announced the preliminary findings of their on-going enquiry (RIKEN 2014). By this time, suggestions of serious misconduct had become an issue, focusing particularly on the work carried out by the STAP cell publications’ lead author, Obokata. Serious problems with several images had been suggested, including ‘cutting and pasting’ (where results had been aligned to suggest the experiments had been carried out at the same time), and duplication (where the same image had been used to show 9

Senior author Teruhiko Wakayama claimed that he gave Obokata these raw results, and may not have been clear about them when he passed them on. Wakayama was also keen to highlight the pressure Obokata was under to complete the manuscripts and get the STAP work published; perhaps her misuse of the images were genuine mistakes.

10 A condition of publishing work in Nature, as of May 2013, is that ‘‘authors are required to make materials, data and associated protocols promptly available to others without undue qualifications’’ (Nature Publishing Group 2014b).

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the results of two apparently different experiments). There were concerns over plagiarism regarding Obokata’s previously published work (such as Obokata et al. 2011), and both image manipulation and plagiarism concerns raised against her PhD thesis.11 RIKEN, in their statement, based on the above, alleged that Obokata had committed misconduct, and noted that the authors accept that inconsistencies in their papers had been damaging for credibility (Research Paper Investigative Committee 2014). Several of the authors had also been considering withdrawing the articles. In response to the requests of the RIKEN committee, author Hitoshi Niwa began his own independent attempts to replicate the STAP cell methods. Despite these apparent misconducts however, Obokata and Vacanti in particular were keen to stress that although there may be some issues with the publications, STAP cells were real and the methods still valid. Apparently for clarity, Vacanti posted a more detailed protocol online; more than a dozen attempts to replicate the work had been reported to date, and none were successful. Influential research laboratory leader Kenneth Ka-Ho Lee (at the Chinese University of Hong Kong) was head of one such group attempting to replicate the work. Lee produced a report for publication in Nature (Brief Communications Arising section) regarding his unsuccessful attempts to reproduce Obokata’s work, however his report was rejected. Continuing his work to establish what had occurred in the laboratory regarding STAP cells, Wakayama announced that he had asked an independent laboratory to carry out a genetic analysis of cells created by Obokata, which she claimed to be STAP cells; Wakayama also carried out some analysis in his own laboratory. In a report which appeared in Nature News on 27 March 2014, Wakayama noted that the cells Obokata had claimed she had created using the STAP cell method were in fact different to those cells Wakayama had given Obokata to test the method.12 On 1 April, RIKEN announced more complete findings of their investigation: this essentially blamed Haruko Obokata for almost the entire controversy. RIKEN announced that they had found Obokata guilty of two incidents of ‘‘research misconduct’’. She ‘‘acted in a manner that can by no means be permitted’’ when she manipulated the data from different experiments. This was alleged to have been carried out to improve the appearance of results. RIKEN also remarked that the poor quality of Obokata’s laboratory notes would make it difficult for anyone else to accurately understand her experiments. Other co-authors of the paper (in particular Wakayama, Sasai, and Niwa) were also deemed ‘‘responsible’’ for not verifying the findings of the work, and for neglecting their mentoring responsibilities towards the young Obokata (Research Paper Investigative Committee 2014). Despite never having interviewed Sasai as part of their review, the RIKEN committee also claimed 11

In early October 2014, The Wall Street Journal reported that Waseda University, where Obokata obtained her doctorate, had threatened to strip Obokata of her PhD; she has been given twelve months to correct her thesis under supervision. Only if she fulfils the university’s requirements will Obokata be allowed to retain her qualification (Martin 2014). 12

In the middle of June 2014, Nature News published a report highlighting exactly how the cells were genetically dissimilar (Cyranoski 2014b). The cells Obokata claimed to be STAP cells had a genetic signature of no other cells at the Center for Developmental Biology at RIKEN. There was a suggestion that the cells presented as STAP cells were probably embryonic stem cells (see also Cyranoski 2014a).

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that as deputy director of the Center for Developmental Biology (CDB), Sasai was motivated by the quest to overshadow the iPS cell work, also carried out in Japan. He was accused by the committee of orchestrating the media reports of Obokata in her laboratory,13 and by being strongly motivated by the potential new grants that STAP could secure. Without consulting Sasai, the committee appeared to make these accusations with few explanations (Cyranoski 2014d). In a press conference on 9 April, Obokata stated that she did not accept the findings of the RIKEN investigation, although admitted ‘‘carelessness’’, ‘‘sloppiness’’, ‘‘immaturity’’ and ‘‘mistakes’’; these however do not affect the conclusion of the papers, Obokata insisted (Otake 2014). Other authors and RIKEN researchers were also under fire, as they retracted papers they had previously published in leading journals.14 Obokata’s co-authors and mentors from Harvard Medical School and Brigham and Women’s Hospital (also in the US), had so far made few comments and had avoided any repercussions. On 30 April, Nature curiously chose to publish a scathing editorial of science policy in Japan. The editorial claimed that few researchers would act on suspicions if they believed their colleagues were involved in misconduct, highlighting that there was no high-level agency in Japan to deal with such allegations (such as the US Office of Research Integrity in America). The editorial also highlighted that sloppiness needed to be distinguished from deception. For the field, this appeared to be an effort to shift blame onto the authors of the STAP publications, and suggested little responsibility would be taken by Nature. A week later, RIKEN announced that they would reject any request by Obokata for re-examination of her work, and that they recommended the retraction of both publications. Such calls were reiterated by many in the scientific community, particularly following F1000’s publication of Lee’s failed replication attempts on 8 May. On 28 May, Obokata agreed to the retraction of the Nature letter; this appeared as somewhat surprising news to some, as it was the research article that was the most concerning for its mistakes. Obokata reiterated that she was poorly mentored, particularly by Wakayama. On 3 June, Obokata also agreed to the retraction of the article; this was relatively unexpected, as Obokata had continued to state that despite any issues with image manipulation, the methods and results of the paper were correct. It may have been that since Obokata’s supporter Vacanti requested the retraction of the article on 30 May, Obokata believed there was no other option. Alongside the internal RIKEN investigation, an external committee had also been called. On 12 June, this committee issued their report. Whilst acknowledging Obokata’s faults, the external committee laid a significant amount of blame at the 13

In Japan, Obokata’s overnight celebrity status was aided by her quirks, again separating her from the stereotypical middle-aged male scientist. For example, she had placed ‘Moomin’ cartoon stickers on her laboratory equipment, and claimed that she didn’t often wear a laboratory coat, but a traditional Japanese apron made for her by her grandmother. 14

This included the head of the RIKEN investigation committee, Shunsuke Ishii. Ishii was suspected of splicing images in figures in a very similar fashion to Obokata; Ishii claimed that this method was considered acceptable ten years ago (when the paper in question was published). Ishii resigned on 25 April, stating that his position would complicate the current investigation of Obokata’s work.

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feet of Sasai and Wakayama for not checking Obokata’s work. The pressure for laboratories at RIKEN to surpass other significant work (such as the development of iPS cells) from Japan in particular, resulted in premature publication and a significant pressure to publish breakthrough work. Poor supervision and oversight was observed through the chain of command at the CDB at RIKEN, and the external committee stated that all involved should face ‘‘severe disciplinary measures’’ (Normile 2014a). The scientific community were also concerned however that despite all of the problems revealed regarding the STAP work in the 4 months since its original publication, Nature had not seen fit to editorially retract the article or letter. The letter and article were officially retracted on 2 July. There was still no comment made from Brigham and Women’s Hospital or Harvard Medical School. Following this, stem cell researcher and blogger Paul Knoepfler published an interview he had with a Nature spokesperson, specifically concerning the editorial and review practices at Nature. The anonymous spokesperson confirmed that the Nature Publishing Group were not members of the Committee for Publication Ethics (COPE), since they believed that the group was focused more on clinical, rather than basic, research (Chair of COPE, Dr Virginia Barbour, commented that COPE has 9000 member journals, and membership is no longer predominantly clinical). Knoepfler also asked about the way Nature papers are checked for plagiarism and image manipulation. Nature use software to check for plagiarism (which, in the case of Obokata et al. 2014b was inadequate), and check a selection of papers for image manipulation (image duplication would be far more complicated to check for, and so far is not carried out on every submission by any journal). Obokata’s manuscripts were not amongst those selected for image analysis (Knoepfler 2014). Had they checked, they may have identified issues with the manuscripts that were later picked-up on PubPeer. Chief editor of the European Molecular Biology Organization journal (EMBO), Bernd Pulverer, requested that image analyst Jana Christopher check the STAP paper, not making Christopher aware of the situation. Christopher identified the spliced gel (considered misconduct by RIKEN), duplicated images (possibly a genuine mistake), and a composite image of cell colonies, possibly created to save space. EMBO and the Journal of Cell Biology have stated that such ‘aberrations’ are found in around 20 % of their submitted manuscripts. This is usually down to naı¨ve manipulation however, often to create neater figures, and without any intention to deceive. Only around 1 % of these papers have their acceptance revoked (Cyranoski 2014c). Defending his journal, Nature editor-in-chief Philip Campbell concluded that neither editors nor reviewers could have been expected to identify the fatal problems that became apparent with the STAP papers (Cyranoski 2014c); this said, it was the observations of ‘lay’ reviewers that identified initial issues with Obokata et al. (2014a, b. Observation of what may initially appear to be minor mistakes can reveal more serious problems with manuscripts. In reply to the accusations that Nature was too keen to publish Obokata et al.’s work without verification, Campbell insists that ‘‘Nature did not let down its guard’’ (Cyranoski 2014c). The reports that STAP cells were not created using the methods described have not only had negative outcomes for Obokata. On 4 August 2014, Yoshiki Sasai committed suicide at the RIKEN centre. In one of apparently several suicide notes,

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Sasai requested that Obokata replicate the STAP cell work (The Japan Times 2014b). Although the RIKEN investigations had cleared Sasai of any direct involvement, he was severely criticised for failing to adequately supervise the writing of the STAP papers, and was reported to have been seeking treatment for depression since March 2014. In a press conference on 12 August, the Sasai family’s lawyer, Kazuhiro Nakamura, summarised the note Sasai had left for his family. It stated that he was ‘‘worn out by the unjust bashing in the mass media and the responsibility he felt towards RIKEN and his laboratory’’. The Japanese tabloid newspapers had written stories based on Sasai’s personal life (such as Shukan Bunshun, that had linked Sasai’s involvement in the STAP cell project to a previous personal relationship he had at university). Sasai was also very troubled by the announcement that the CDB would be dismantled, as recommended by the RIKEN committee. This would result in downsizing, renaming, and a re-launch later in 2014 (Normile 2014b). RIKEN announced at the end of August 2014 that 22 separate attempts had been made to replicate the STAP protocol, however none had been successful. The unsuccessful results of Niwa’s attempts were announced on 27 August. Niwa suggested that he had seen green fluorescence as expected from the green fluorescent protein (GFP) the cells were labeled with, however concluded that this was the autofluorescence observed during cell death, and not indicative of a successful result (Cyranoski 2014e). Following Niwa’s announcement, Vacanti provided a revised protocol online (Vacanti & Kojima 2014). The protocol, dated 3 September 2014, suggested that the addition of ATP was required to the cells, in addition to the physical stress and acid treatment. In late September 2014, Takaho Endo, a researcher at RIKEN’s Center for Integrative Medical Science, published a paper in Genes to Cells which described how a technique utilising RNA sequencing could be used to identify cell types (Endo 2014).15 To demonstrate the use of this technique, Endo bravely selected to use the cells Obokata had claimed to be STAP cells in her recently retracted research article. As well as demonstrating that RNA sequencing could differentiate between different cell types, Endo reported that the apparent STAP cells had Trisomy 8, which is known to be lethal at the embryonic stage (i.e. cells with this mutation are not capable of contributing to embryonic development); this is a well-known trait of some embryonic stem cell lines, and the most common chromosomal abnormality to occur in mice. Although this is not stated by Endo, the identification of Trisomy 8 in Obokata’s STAP cells strongly suggests that these cells did not contribute to the development of the live mice as reported. Prior Publication Attempts On 10 September 2014 further details of the Science reviewers comments were made available online (Retraction Watch 2014).16 An unnamed senior editor at Science returned three reviewers comments to Obokata on 21 August 2012 (whilst Obokata 15 The RNA sequencing (RNA-seq) Endo describes looks specifically for expression levels of certain genes and the frequency of alleles in the cell population tested. 16

The manuscript originally sent to Science had Wakayama as a co-author, but not Sasai.

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was at her previous post, at the Brigham and Women’s Hospital/Harvard Medical School). The Science senior editor rejected Obokata’s manuscript (then titled ‘‘Stress altered somatic cells capable of forming an embryo’’), since despite the prospect of Obokata addressing many of the specific criticisms raised in a revised manuscript, the general negative reviews of the work are such that Science would not publish this work anyway. More significantly, the comments made by the three anonymous Science reviewers cover some of the significant issues raised by others after the revised manuscript had been accepted and published by Nature in January 2014. This includes the concerns about splicing lanes from different gels to appear that they were from the same gel. In the relatively lengthy reviews, there are also requests for more detail in the methods and protocols. Such comments coincided with the initial reviewers comments from Nature, as reported in the RIKEN report. As Obokata stated to the RIKEN committee however, she did not carefully consider the comments of the Science reviewers (possibly since the senior editor had stated that Science would not be interested in publishing a revised manuscript anyway). Apparently in relation to this, Science were also able to publish some email correspondence between Obokata and Nature regarding the letter and original research article submitted to them in March 2013. On 4 April 2013, Nature initially rejected the papers. Unlike Science however, Nature stated that if ‘further experimental data allow you to address these criticisms, we would be happy to look at a revised manuscript’ (Vogel and Normile 2014); by 20 December 2013, Nature were clearly satisfied that Obokata et al. had addressed the criticisms made by its reviewers, and accepted the letter and article for publication.

Trust, Replication, and STAP Cells Trust For the purposes of this paper, the following definition of trust is useful: ‘‘trust means deferring with comfort and confidence to others, about something beyond our knowledge or power, in ways that can potentially harm us’’ (Whyte and Crease 2010, p 412). Research in recent decades has become particularly collaborative, with an emphasis on multidisciplinary studies. From the earliest stages, trust is a significant issue. Researchers rely on each other through every stage of the research process: for example, when informally discussing ideas for a funding proposal; giving fair critique when reviewing a research proposal; trusting that each collaborator will contribute as expected to a research project; advising collaborators to the best of their ability; and so on. There is a lot of trust extended and expected prior to publication of results. As well as trusting each other’s laboratory work, researchers need to trust each other as they review colleague’s work. The peer-review process, considered as important as replication by some,17 is now an established part of 17 For example, according to the University of Tromsø Pro Rector for Research and Development Curt Rice, peer-review is a ‘‘sacred cow’’ of scientific research (Rice 2013).

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publishing. Anonymity during the peer-review process encourages virtues such as openness, truthfulness, and trust (from authors and reviewers). Replication Replication has long been considered an essential part of scientific progress.18 There are several uses for replication in biological science research, including increased confidence in a result, reliability of results, quality of work, and, to an extent, the reputation of the researchers, institutes and universities where the research is being carried out. For these reasons, replication has been considered by some to be the best defence against fraud.19 There are problems with undertaking replications however, despite the acceptance of the theory. Firstly, there is often a lack of methodological detail recorded in scientific publications. For a laboratory attempting to repeat a study based only on published articles, small nuances of method may not be detailed, causing the repeating laboratory to potentially fail to obtain the same results.20 Secondly, there is a lack of available resources. Repeating a study may require significant time and resources (such as use of expensive equipment), which can not always be spared or obtained. This may be particularly true of certain medical trials for example, where specific groups of patients may be required to replicate a study. It has also been demonstrated that often ‘interesting’ results, not those that are repeatable, are published.21 Repeating the work of another laboratory or researcher may also be considered a challenge to others’ integrity or skill, capable of causing animosity in the scientific community (Broad and Wade 1982; Koole and Lakens 2012). Lastly, there is a lack of motivation for researchers to carry out replication studies. Researchers are motivated by both scientific and personal goals; progress in science is about creating new knowledge (and not about deliberately embarking on work to 18

For example, see Zuckerman (1984), Hardwig (1991), Jasny et al. (2011), Crocker and Cooper (2011), Koole and Lakens (2012), Chambers and Sumner (2012).

19 For example, also considering peer-review and refereeing (unfortunately without highlighting the difference between the two), Broad and Wade (1982) suggest that replication is the third line of defence and ‘‘most exacting test’’ for potentially fraudulent claims (p 17, 62). 20

In some cases, deliberate omissions may be made in order to prevent replication of an experiment. For example, Resnik (1998) highlights the case of chemists Stanley Pons and Martin Fleischmann; when they believed they had found a method for successful cold fusion, Pons and Fleischmann had financial incentives for being vague about their methods. Robert Proctor (2008) has also pointed out the importance of ‘institutional amnesia’ for those working in the military, law, or government for example (p 10). Peter Galison (2008) goes a step further, suggesting that secrecy may even be a threat to democracy. Stepping back from such ambitious claims, Frederick Grinnell (1992) has suggested that science does not necessarily require secrecy, but ‘discretion’; secrecy would likely bring scientific progress to a standstill, but discretion is seen as necessary. Priority is of particular importance in the current scientific climate, and appropriate discretion can be the difference between getting funded or not (p 123). 21 Writing for a special issue of Science, Jennifer Couzin-Frankel has described a situation whereby a researcher was struggling to replicate the findings of a study published in a highly-regarded journal. The researcher, whilst at a conference, had the opportunity to talk to the author of the paper, and quizzed him about the experiment. Nonchalantly, the author replied that they too had repeated the experiment many times, but the described result only occurred once. Since that was the interesting result, this is the one published (Couzin-Frankel 2013).

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demonstrate that someone else’s study was wrong). As previously mentioned, replication studies also utilise resources that could instead be used to undertake new, original work.

Trust and Replication in the STAP Cell Controversy With regard to trust, there are several issues in this story that have come to light. Firstly, regarding Obokata et al. (2014b) on the article and letter. Wakayama had carried out the work on mouse development, specifically referred to in the letter. Since there has been controversy regarding the use of the raw data (i.e. the same image used to demonstrate the results of two different experiments), Wakayama has suggested that perhaps Obokata did not understand exactly what the raw data was, or that he himself had not explained it clearly enough when handing the raw results over. Wakayama clearly trusted that Obokata then was going to use the data appropriately in the preparation of her manuscripts, whilst Obokata trusted that Wakayama had given her the results of the experiments he had carried out. This is a standard procedure in laboratories, where often several individuals may be involved in different aspects of a single project. In this case, all of the co-authors on the letter and original research articles believed that Obokata was putting together two honest manuscripts. The trust placed in Obokata by her co-authors may also explain why none of these individuals had seen or corrected the faults in the papers prior to submission. It may have been the case however that few co-authors knew what the Cell, Science, or Nature reviewers comments were, since there is usually only one corresponding author listed (in this case, Obokata). It may be difficult to carry out, and imply that researchers can not trust one another, but one suggestion from the STAP cell case to prevent such an occurrence from happening again is that co-authors and collaborators should carry out verification tests prior to submission. For example, Wakayama requested that an independent company carry out genetic testing of STAP cells and other embryonic cells from the CDB at RIKEN, that demonstrated a discrepancy in the cells Wakayama gave to Obokata, and the cells Obokata returned to Wakayama, claiming to have successfully converted them to STAP cells; arguably there was certainly time for this to have been achieved following Nature’s first rejection of the article, and its resubmission. In fact, if the results had been consistent, it would have helped to substantiate the STAP claim. This leads us to consider another, more general aspect of modern science: the pressure to publish. It has been suggested that Obokata (and presumably the other RIKEN authors) were under significant pressure to publish work to rival the iPS cell breakthroughs made by Shinya Yamanaka: pressure to maintain Japan’s place at the pinnacle of stem cell and regenerative research. The pressure to publish is a significant motivator for researchers (particularly early career scientists like Obokata), and this would have certainly had an effect on the manuscripts Obokata submitted. In addition, it may explain why Obokata and her co-authors apparently took little notice of the Science reviewers comments (since Science had already stated that they would not be interested in publishing a revised manuscript). At the

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moment however, although some notes of the Cell review and the Science review have been made available to the public, there has been no such release of detailed Nature reviewers comments; this said however, on 11 September 2014, Science published an online news story covering email correspondence between Nature and Obokata regarding the two manuscripts that were later revised, accepted, and published (Vogel and Normile 2014). Wakayama here states that he was not sent the reviewers comments personally, however Obokata shared them with him. Wakayama claims that he made revision suggestions to Obokata regarding his contribution (i.e., creating the chimeric mice), but did not know whether a revised manuscript was sent for further review. Perhaps, Wakayama suggests, all co-authors should be sent copies of reviewer comments on submitted manuscripts, regardless of whether the section or results they contributed required revision. This apparent lack of communication between co-authors (possibly based on the trust they had for one another), was highlighted by the external RIKEN committee, who suggested that there were significant issues with the chain of command, and that all should face disciplinary action. Such a response from the committee suggested that any trust RIKEN researchers had in each other was misplaced. It is possibly much easier to suggest this with hindsight however, but is it reasonable to suggest that all of the co-authors on both papers should have been explicitly informed and involved at each step of the publication process? Does this instruction imply that all co-authors must see reviewers comments (as suggested by Wakayama), and contribute to revisions? This is not the usual process manuscripts go through prior to submission and acceptance; why would the RIKEN researchers have expected their work to be any different? A further, more general comment made in a Nature online report in April 2014, suggests that there is no culture of reporting scientific misconduct in Japan. Although the aims of this Nature piece need to be considered carefully, the lack of misconduct reporting in Japan may have a role in the scientific culture of the country. It also suggests however that the lack of a Japanese equivalent of the US Office of Research Integrity may be because there is less severe misconduct to report. Although the current STAP cell controversy has highlighted an issue which occurs in all scientific communities, few other reports of misconduct in Japanese research have been reported previously. I am not suggesting that there is no misconduct in Japanese science, and that the current controversy is an isolated event, but is lack of reporting due to the lack of misconduct? This is unlikely. There is a further distinction that needs to be made between sloppiness and deception; where there is significant pressure to publish, sloppiness can creep into work, regardless of the stature of those carrying it out. This is not necessarily deliberate deception however. As Obokata has previously claimed, there may be errors in the publications (‘sloppiness’), however STAP cells are real (no deception was intended). Replication of the STAP cell protocols, several months following publication (and now retraction) of the article and letter, has not yet been successfully achieved. With the ability to use the internet to disseminate reports and comments, the scientific community was alerted to several replication attempts within days of original publication. In addition, after only a week post-publication, reports

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appeared that replication attempts had been unsuccessful. Initially, there was a reasonable explanation for this (that did not include the concern that STAP cells did not exist): that there was possibly a lack of methodological detail in the article.22 Although this was a supposedly simple method, other laboratories were not succeeding in producing the desired results. In March 2014, Charles Vacanti posted more detailed protocols online, presumably in an effort to demonstrate that there was a standard method for creating STAP cells, and that it was relatively simple.23 Vacanti’s protocol should also have allowed others to replicate the method exactly as had been carried out at RIKEN. Despite this, March 2014 reports suggested that a dozen independent groups had attempted to replicate STAP cell creation and had all failed. Interestingly, the results of influential cell biologist Prof Ken Lee’s replication attempts were rejected for publication. Lee initially published his preliminary results on ResearchGate, a website that invites review of publications using a feature called ‘Open Review’. As part of the Open Review for the STAP article (Obokata et al. 2014a), Lee and his laboratory made several attempts to replicate the STAP protocol from the methods given in the article, and, later, using the more detailed protocols released by Vacanti. Lee put together a report on his replication work, which was submitted to Nature’s Brief Communications Arising section; an online only resource that seeks to include ‘‘exceptionally interesting or important scientific comments and clarifications on original research papers or other peer-reviewed material published in Nature’’ (Nature Publishing Group 2014a). The usual procedure is that, if the editors see fit, the communication is passed to the original authors of the article for comment. In Lee’s case, Senior Editor at Nature, Francesca Cesari, declined to publish Lee’s communication [since the report did not ‘‘challenge key data or conclusions’’ Cesari (2014)], and presumably it was not sent to Obokata et al. for consideration. Fortunately, as well as submitting brief updates on the ResearchGate website (as noted in ResearchGate 2014), Lee and his laboratory were able to get a report published in the journal F1000Research, an open science journal with invited, open peer review and discussion (Tang et al. 2014). It has been made relatively clear then that the scientific community, unsurprisingly, wanted to replicate Obokata et al.’s work as quickly as possible. Although Koole and Lakens (2012) suggest that carrying out replication studies may be considered a ‘challenge’ to the original laboratory and its work, I do not believe that this was the case for STAP cells. I believe that the stem cell community were genuinely intrigued and excited at the prospect of inducing dedifferentiation by such a relatively simple method. The swift exercises to replicate the STAP cell protocols were not, I suggest, to demonstrate that Obokata’s work was faulty. Instead, it was to reproduce her technique and to be able to use STAP cells as a tool in their own 22 There are often few methodological details noted in original research articles, particularly in high impact journals such as Nature, where space is often at a premium. Small nuances of protocols may not be reported explicitly, and this is (currently) normal practice. Usually, if contacted, a laboratory is willing to share more detailed protocols or offer a visit, to allow others to replicate methods. 23 It perhaps took almost two months for Vacanti to publicise the more detailed protocol, as an initial interview on 2 February suggested that the original research article (Obokata et al. 2014b) was essentially a ‘how-to’ of creating STAP cells.

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laboratories around the world, as well as develop the technique to include human cells. If this were possible, there would be much that could be learned for both basic biology and for applied, particularly for therapeutic and regenerative medicine. This is justified by the first reactions of researchers: surprised at the simplicity, but hoping that this was a valid method, replication studies began prior to any comments arising about sloppiness or potential misconduct. The scientific community’s first response was to recreate Obokata’s methods, allowing STAP cells to become an invaluable research tool in laboratories around the world. Replication studies however did not seem to concern the Japanese press. Japanese media initially focused on their new face of science. Once concerns regarding the presentation of figures in the article and letter began to emerge however, the media were far more interested in the potential misconduct that had arisen, with little or no mention of replication studies. I propose that there are two possibilities for this. Firstly, the popular media has a general misunderstanding of how science is carried out and verified, including replication of protocols. The press therefore did not realise the scientific impact of so many laboratories reporting that they could not reproduce Obokata’s results. The second possibility is simply that the misconduct story was far more interesting. As demonstrated above, there were developments and new reports and comments on this story on (at least) a weekly basis, providing plenty for the Japanese media to report on. Following rumours of Obokata being chased by reporters and photographers [and, on at least one occasion, coming to harm following this (The Japan Times 2014a)], I believe that this latter is more likely; the misconduct story and Obokata’s fall from grace (where the media had placed her in the first place), is a far more interesting angle to report from, than from a science studies angle considering the importance of replication, and, of course, whether STAP cells actually exist.

Conclusions In February 2004, Hwang Woo Suk announced that his laboratory had generated human stem cell lines from cloned embryos, and published two papers demonstrating the work. Two years later, these papers were retracted, and Hwang’s laboratory work shown to be fraudulent. Scientific integrity and authorship came under scrutiny. Nature and other prominent journals then promised that the review system would be more careful from now on. The publishing establishment genuinely appeared to alter their practices in the light of Hwang’s fraud, and when in 2007, Shoukhrat Mitalipov claimed that his laboratory had cloned embryonic stem cell lines from monkeys, Nature requested an independent validation of the results. Once received, Mitalipov’s work was published alongside the verification. Many in the scientific community agreed and applauded Nature’s stringency. Why then, in 2014, were Obokata’s manuscripts not put through the same rigorous testing as Mitalipov’s? It would be difficult to suggest that the protocols of large numbers of papers are outsourced and replication studies carried out before publication— such external verification is not feasible. This would take time and resources away from laboratories that have their own interests to pursue. (This is demonstrated by

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the post-publication replication efforts of Ken Lee’s laboratory; after several attempts, using different protocols, Lee had to stop his unsuccessful attempts to create STAP cells and continue working on his own project.) So then, is Obokata a victim of trust and replication in science and its new culture (regardless of whether fraud was committed or not)? What roles have trust and replication had in the case of STAP cells? With regard to trust, it has been suggested that Obokata became generally untrustworthy to those collaborating with her. For example, some authors now claim that they had not seen Obokata’s data, or that they can not replicate the methods themselves. In a scientific culture that presumes honesty, truthfulness, openness, and integrity, Obokata’s perceived misconduct goes against how her peers expected her to act. As well as such career-driven motivations, researchers have personal motivations, which, due to the nature of their work, often include strong intellectual curiosity and ‘emotional drive’ (Stevenson and Byerly 1995). Other virtues such as honesty and integrity are usually considered standard qualities for scientific researchers, and training for early career researchers often reflects this (Shapin 2008). This has led some to suggest that to act fraudulently is rare, and ‘not normal’ behaviour, since a regular scientific researcher would not believe she/he could get away with misconduct (Resnik 1998). With regards to replication, I have suggested that experiments are usually repeated in the original laboratory by the original laboratory workers several times prior to publication. Obokata has stated that she created STAP cells many, many times. Several researchers appear to have attempted the methods outlined in Obokata’s papers to create their own STAP cells. Many laboratories began this replication soon after the paper was published, although, as expected, it was difficult to publish these results. Once the misconduct accusation had been made, researchers from other laboratories who had been attempting to replicate the work began reporting their difficulties. Eventually, blame shifted to Obokata almost exclusively. Despite Broad and Wade (1982) suggesting that in fact replication is ‘‘a theoretical construct dreamed up by philosophers and sociologists of science’’ (p 77), I believe that researchers generally accept the need for replication in principle. As highlighted by Karl Popper’s views on practical science, the theory considered most correct at any one time is the one that has been tested the most; experimentation can only disprove, not prove theories, and at all times researchers work to test what is already ‘known’. If we not only include new results, but new methods in this theory, only methods that work will continue to be used (and replicated). Those that do not will be consigned to history (Grinnell 1992). New methods therefore have the opportunity to become new tools for research, moving from an interesting and useful result into the realm of typically used methods; particularly successful methods and tools will consequently be ‘black boxed’ as they become more commonly utilised. This essay has also raised other questions regarding trust and replication in science, such as whether all co-authors should be involved in the publication process. Publications are currently essential for a researcher’s career, since they have impacts on grants awarded, positions offered, and, essentially, the livelihood of researchers. Generally, journals are particularly unwilling to publish replication studies. Such studies are regarded as dull (Chambers and Sumner 2012), not

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newsworthy, and a waste of space. If publication is a significant motivator for research, then replication studies are significantly disincentivised. The answer to the above question then should be yes—all co-authors should receive and be able to comment on reviewer comments, and sign agreements to publish. The downside of this of course is that publication will take longer, but is this a relatively small price to pay for better science communication? Unfortunately, such actions however also suggest a lack of trust (and, to an extent, responsibility) between researchers. Why should journals not be able to trust an author to share his or her reviewer comments, and corrected manuscripts with their co-workers? There is evidence that this does not happen—papers have been retracted by authors since they claimed they were not involved in the publication process at all (for example, see Huss et al. 200824). Such a step appears to admit that there is a shroud of mistrust in scientific culture; should this be addressed explicitly (for example, by making all authors equally responsible for publications), or more subtly managed? It is possible that not enough change will occur from the latter. The aim of this essay was to show the ongoing relationship between trust and replication with specific reference to the STAP cell controversy. Although trust can exist prior to (and in the absence of) replication, failure to replicate can be driven by lack of trust. In this case study, we can see that the expected behaviour of scientists, trust, and replication all had a role in the case against Dr Haruko Obokata; there is perhaps too much trust in modern biological science, and not enough replication. Acknowledgments A shorter version of this paper was presented at the Centre for Ethics, Law, and the Life Sciences (CELLS), Durham University, in June 2014. I would like to thank CELLS members, and in particular Prof A-H Maehle, for their comments, and to the anonymous reviewers for their helpful suggestions.

References Broad, W., & Wade, N. (1982). Betrayers of the truth: A fraud and deceit in the halls of science. New York: Simon and Schuster. Cesari, F. (2014). Letter to professor Lee. http://www.researchgate.net/publication/259984904_Stimulustriggered_fate_conversion_of_somatic_cells_into_pluripotency/reviews/103. Accessed 4 Sept 2014. Chambers, C., & Sumner, P. (2012). Replication is the only solution to scientific fraud. The guardian. http://www.theguardian.com/commentisfree/2012/sep/14/solution-scientific-fraud-replication. Accessed 8 April 2014. Chen, Z.-X., & Riggs, A. D. (2011). DNA methylation and demethylation in Mammals. Journal of Biological Chemistry, 286, 18347–18353. Couzin-Frankel, J. (2013). The power of negative thinking. Science, 342, 68–69. Crocker, J., & Cooper, M. L. (2011). Addressing scientific fraud. Science, 332, 1182. Cyranoski, D. (2014a, 3 June). Genetic tests suggest STAP stem cells ‘never existed’. Nature. http:// blogs.nature.com/news/2014/06/genetic-tests-suggest-stap-stem-cells-never-existed.html. Accessed 1 Oct 2014. Cyranoski, D. (2014b, 17 Jun). Gene tests suggest acid-bath stem cells never existed. Nature. http://www. nature.com/news/gene-tests-suggest-acid-bath-stem-cells-never-existed-1.15425. Accessed 1 Oct 2014.

24 The retraction notice in Der Pathologe for Huss, Xiao, and Hiemberg states that neither Heimberg nor Xiao had given their consent for publication, ‘‘whose existence they were not aware of’’. Huss et al. (2008, p 289).

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C. Lancaster Cyranoski, D. (2014c, 3 Jul). Research integrity: Cell-induced stress. Nature. http://www.nature.com/ news/research-integrity-cell-induced-stress-1.15507. Accessed 1 Oct 2014. Cyranoski, D. (2014d, 13 Aug). Stem-cell pioneer blamed media ‘bashing’ in suicide note. Nature. http:// www.nature.com/news/stem-cell-pioneer-blamed-media-bashing-in-suicide-note-1.15715. Accessed 1 Oct 2014. Cyranoski, D. (2014e, 12 Sep). STAP co-author offers yet another recipe for stem cells. Nature. http://blogs.nature.com/news/2014/09/stap-co-author-offers-yet-another-recipe-for-stem-cells.html. Accessed 18 Oct 2014. Endo, T. A. (2014). Quality control method for RNA-seq using single nucleotide polymorphism allele frequency. Genes to Cells,. doi:10.1111/gtc.12178. Fang, F. C., Steen, R. G., & Casadevall, A. (2012). Misconduct accounts for the majority of retracted scientific publications. PNAS, 109(42), 17028–17033. Galison, P. (2008). Removing knowledge: The logic of modern censorship. In R. N. Proctor & L. Schiebinger (Eds.), Agnotology: The making and unmaking of ignorance (pp. 37–54). California: Stanford University Press. Grinnell, F. (1992). The scientific attitude (2nd ed.). London: The Guilford Press. Hardwig, J. (1991). The role of trust in knowledge. The Journal of Philosophy, 88(12), 693–708. House of Lords. (2000). Science and technology—Third report. http://www.publications.parliament.uk/ pa/ld199900/ldselect/ldsctech/38/3801.htm. Accessed 1 Jan 2015. Huss, R., Xiao, X., & Heimberg, H. (2008). Retraction: Adult stem cells regenerate the endocrine pancreas and normalize hyperglycaemia and insulin production in diabetic mice. Der Pathologe, 29(2), 289. Jasny, B. R., Chin, G., Chong, L., & Vignieri, S. (2011). Again, and again, and again …. Science, 334, 1225. Knoepfler, P. (2014, 3 July). Interview with Nature on their editorial process in wake of STAP. Knoepfler lab stem cell blog. http://www.ipscell.com/2014/07/interview-with-nature-on-their-editorialprocess-in-wake-of-stap/. Accessed 1 Oct 2014. Koole, S. L., & Lakens, D. (2012). Rewarding replications: A sure and simple way to improve psychological science. Perspectives on Psychological Science, 7, 608–614. Martin, A. (2014). Japanese university may strip stem-cell scientist of doctorate. The Wall Street Journal. http://online.wsj.com/articles/japanese-university-may-strip-stem-cell-scientist-of-doctorate1412683955. Accessed 18 Oct 2014. Martin, A. & Naik, G. (2014). Japanese institute weighs retracting stem-cell studies: Move by Riken comes a day after co-author called for retraction, citing ‘crucial mistakes’. The Wall Street Journal. http://online.wsj.com/articles/SB10001424052702304020104579430600241800532. Accessed 18 Oct 2014. Nature Publishing Group. (2014a). For authors. Brief communications arising. http://www.nature.com/ nature/authors/gta/commsarising.html. Accessed 4 Sept 2014. Nature Publishing Group. (2014b). Availability of data & materials: Authors & referees. http://www. nature.com/authors/policies/availability.html. Accessed 18 Oct 2014. Normile, D. (2014a, 12 Jun). Japanese stem cell debacle could bring down center. Science. http://news. sciencemag.org/asiapacific/2014/06/japanese-stem-cell-debacle-could-bring-down-center. Accessed 18 Oct 2014. Normile, D. (2014b, 27 Aug). In Japan, official effort to replicate STAP stem cells comes up empty. Science. http://news.sciencemag.org/asiapacific/2014/08/japan-official-effort-replicate-stap-stemcells-comes-empty. Accessed 1 Oct 2014. Normile, D., & Vogel, G. (2014). 13 Jun). STAP cells succumb to pressure: Retraction plans for easy stem cell recipe leave scientists wondering how the papers came to be published. Science, 344(6189), 1215–1216. Obokata, H., Kojima, K., Westerman, K., Yamato, M., Okano, T., Tsuneda, S., & Vacanti, C. A. (2011). The potential of stem cells in adult tissues representative of the three germ layers. Tissue Engineering Part A, 17(5–6), 607–615. Obokata, H., Sasai, Y., Niwa, H., Kadota, M., Andrabi, M., Takata, N., et al. (2014a). Bidirectional developmental potential in reprogrammed cells with acquired pluripotency. Nature, 505, 676–680. Obokata, H., Wakayama, T., Sasai, Y., Kojima, K., Vacanti, M. P., Niwa, H., et al. (2014b). Stimulustriggered fate conversion of somatic cells into pluripotency. Nature, 505, 641–647. Otake, T. (2014). Obokata says STAP cell discovery not fabrication, claims RIKEN dissuaded her from giving her side of the story earlier. The Japan Times. http://www.japantimes.co.jp/news/2014/04/09/

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The Acid Test for Biological Science national/obokata-says-stap-cell-discovery-not-fabrication-claims-riken-dissuaded-her-from-givingher-side-of-story-earlier/#.VELYVl5–lI. Accessed 18 Oct 2014. Proctor, R. (2008). Agnotology: A missing term to describe the cultural production of ignorance (and its study). In R. N. Proctor & L. Schiebinger (Eds.), Agnotology: The making and unmaking of ignorance (pp. 1–33). California: Stanford University Press. Research Paper Investigative Committee. (2014). Report on STAP cell research paper investigation. http://www3.riken.jp/stap/e/f1document1.pdf. Accessed 18 Oct 2014. ResearchGate. (2014, Apr 1). Researcher partly validates results of controversial stem cell study. ResearchGate. https://explore.researchgate.net/display/news/2014/04/01/Researcher?partly?validates ?results?of?controversial?stem?cell?study. Accessed 1 Oct 2014. Resnik, D. B. (1998). The ethics of science: An introduction. London: Routledge. Retraction Watch. (2014, Sept 10). ‘‘Truly extraordinary,’’ ‘‘simply not credible,’’ ‘‘suspiciously sharp:’’ A STAP stem cell peer review report revealed. Retraction watch http://retractionwatch.com/2014/ 09/10/truly-extraordinary-simply-not-credible-suspiciously-sharp-a-stap-stem-cell-peer-reviewreport-revealed/. Accessed 12 Sept 2014. Rice, C. (2013, Oct 4). Open access publishing hoax: What science magazine got wrong. The Guardian. http://www.theguardian.com/higher-education-network/blog/2013/oct/04/science-hoax-peerreview-open-access. Accessed 5 Oct 2013. RIKEN. (2014). Interim report on the investigation of the Obokata et al. articles. http://www.riken.jp/en/ pr/press/2014/20140314_1/. Accessed 18 Oct 2014. Shapin, S. (2008). The scientific life: A moral history of a late modern vocation. London: University of Chicago Press. Stevenson, L., & Byerly, H. (1995). The many faces of science: An introduction to scientists, values and society. Colorado: Westview Press. Takahashi, K., & Yamanaka, S. (2006). Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126(4), 663–676. Tang, M. K., Lo, M. L. M., Shi, W. T., Lee, H. S. S., & Lee, K. K. H. (2014). Transient acid treatment cannot induce neonatal somatic cells to become pluripotent stem cells. F1000Research. doi: 10.12688/f1000research.4092.1. http://f1000research.com/articles/3-102/v1#sthash.7XanTt6W.dpuf. Accessed 4 Sept 2014. The Japan Times. (2014a). Obokata injured while fleeing pack of NHK reporters, lawyer says. http:// www.japantimes.co.jp/news/2014/07/24/national/obokata-injured-fleeing-pack-nhk-reporterslawyer-says/#.VELSsF5–lI. Accessed 18 Oct 2014. The Japan Times. (2014b). STAP paper co-author Sasai commits suicide. http://www.japantimes.co.jp/ news/2014/08/05/national/embattled-stap-study-co-author-dies-after-apparent-suicide-bid/#.U_ Srjtq9KSN. Accessed 18 Oct 2014. Thomson, H. (2014, 5 Feb). Extraordinary stem cell method tested in human culture. New Scientist. http:// www.newscientist.com/article/dn25004-extraordinary-stem-cell-method-tested-in-human-tissue.html. Accessed 1 Oct 2014. Vacanti, C. A., & Kojima, K. (2014). Revised STAP cell protocol. 09.03.14. https://research. bwhanesthesia.org/site_assets/51520d191eea6679ce000001/cterm/Revised_STAP_protocol28bcd7e61d02a23624eb590717e241fe.pdf. Accessed 18 Oct 2014. Vogel, G., & Normile, D. (2014). Exclusive: Nature reviewers not persuaded by initial STAP stem cell papers. Science. http://news.sciencemag.org/asiapacific/2014/09/exclusive-nature-reviewers-notpersuaded-initial-stap-stem-cell-papers. Accessed 12 Sept 2014. Whyte, K. P., & Crease, R. (2010). Trust, expertise and the philosophy of science. Synthese, 177(3), 411–425. Zuckerman, H. (1984). Norms and deviant behaviour in science. Science, Technology and Human Values, 9(1), 7–13.

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