RESEARCH ARTICLE
Differentiation of human embryonic stem cells into corneal epithelial progenitor cells under defined conditions Canwei Zhang1,2, Liqun Du1, Kunpeng Pang1, Xinyi Wu1* 1 Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, PR China, 2 The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital of Shandong University, Jinan, Shandong, PR China
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OPEN ACCESS Citation: Zhang C, Du L, Pang K, Wu X (2017) Differentiation of human embryonic stem cells into corneal epithelial progenitor cells under defined conditions. PLoS ONE 12(8): e0183303. https:// doi.org/10.1371/journal.pone.0183303 Editor: Wei Li, Xiamen University, CHINA Received: December 18, 2016 Accepted: August 2, 2017 Published: August 15, 2017 Copyright: © 2017 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: This work was supported by the National Natural Science Fund of China (No. 81271716; https://isisn.nsfc.gov.cn/egrantindex/funcindex/ prjsearch-list) and Key research and development program of Shandong Province (No. 2015GGE27292; http://www.sdstc.gov.cn/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
* [email protected].
Abstract The development of cell-based therapies using stem cells represents a significant breakthrough in the treatment of limbal stem cell deficiency (LSCD). The aim of this study was to develop a novel protocol to differentiate human embryonic stem cells (hESCs) into corneal epithelial progenitor cells (CEPCs), with similar features to primary cultured human limbal stem cells (LSCs), using a medium composed of DMEM/F12 and defined keratinocyte serum-free medium (KSFM) (1:1) under different carbon dioxide (CO2) levels in culture. The differentiated cells exhibited a similar morphology to limbal stem cells under 5%, 7%, and 9% CO2 and expressed the LSC markers ABCG-2 and p63; however, CK14 was only expressed in the cells cultured under 7% and 9% CO2. Quantitative reverse-transcription polymerase chain reaction (RT-PCR) analysis indicated that the ABCG2, p63, and CK14 levels in the 7% CO2 and 9% CO2 groups were higher than those in the 5% CO2 group and in undifferentiated hESCs (p
Differentiation of human embryonic stem cells into corneal epithelial progenitor cells under defined conditions Canwei Zhang1,2, Liqun Du1, Kunpeng Pang1, Xinyi Wu1* 1 Department of Ophthalmology, Qilu Hospital of Shandong University, Jinan, Shandong, PR China, 2 The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital of Shandong University, Jinan, Shandong, PR China
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OPEN ACCESS Citation: Zhang C, Du L, Pang K, Wu X (2017) Differentiation of human embryonic stem cells into corneal epithelial progenitor cells under defined conditions. PLoS ONE 12(8): e0183303. https:// doi.org/10.1371/journal.pone.0183303 Editor: Wei Li, Xiamen University, CHINA Received: December 18, 2016 Accepted: August 2, 2017 Published: August 15, 2017 Copyright: © 2017 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: This work was supported by the National Natural Science Fund of China (No. 81271716; https://isisn.nsfc.gov.cn/egrantindex/funcindex/ prjsearch-list) and Key research and development program of Shandong Province (No. 2015GGE27292; http://www.sdstc.gov.cn/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
* [email protected].
Abstract The development of cell-based therapies using stem cells represents a significant breakthrough in the treatment of limbal stem cell deficiency (LSCD). The aim of this study was to develop a novel protocol to differentiate human embryonic stem cells (hESCs) into corneal epithelial progenitor cells (CEPCs), with similar features to primary cultured human limbal stem cells (LSCs), using a medium composed of DMEM/F12 and defined keratinocyte serum-free medium (KSFM) (1:1) under different carbon dioxide (CO2) levels in culture. The differentiated cells exhibited a similar morphology to limbal stem cells under 5%, 7%, and 9% CO2 and expressed the LSC markers ABCG-2 and p63; however, CK14 was only expressed in the cells cultured under 7% and 9% CO2. Quantitative reverse-transcription polymerase chain reaction (RT-PCR) analysis indicated that the ABCG2, p63, and CK14 levels in the 7% CO2 and 9% CO2 groups were higher than those in the 5% CO2 group and in undifferentiated hESCs (p
