TRANSLATIONAL AND CLINICAL RESEARCH 1‐ GMP‐compliant Stem Cell Facili‐ ty, Brain and Spinal Cord Injury Research Center, Tehran University of Medical Sciences, Tehran, Iran; 2‐ Endocrinology and Metabolism Research Center, Tehran University of Medical Sciences, Tehran, Iran; 3‐ GMP Cellular Therapy Facility, Chronic Diseases Research Center, Tehran University of Medical Sci‐ ences, Tehran, Iran
Cell Manufacturing for Clinical Applications BABAK ARJMAND1,2, HAMID REZA AGHAYAN2,3 Key words. AD‐MSCs • Cell manufacturing • GCP • GMP STEM CELLS 2014; 00:000–000
Corresponding author: Hamid Reza Aghayan, Endocrinology and Me‐ tabolism Research Center and GMP Cellular Therapy Facility, Chronic Diseases Research Center, Tehran University of Medical Sciences, Tehran, Iran. E‐Mail: [email protected] Received February 22, 2014; ac‐ cepted for publication March 16, 2014; available online without sub‐ scription through the open access option. ©AlphaMed Press 1066‐5099/2014/$30.00/0 This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typeset‐ ting, pagination and proofreading process which may lead to differ‐ ences between this version and the Version of Record. Please cite this article as doi: 10.1002/stem.1751 To the editor: The study of adipose tissue derived MSCs (AD‐MSCs) injection for osteoarthritis by Jo et al., was fascinating to us [1]. It is a promising study with good clinical trial design that contains valuable clinical information. In contrast to clinical data, cell manufacturing process has been briefly described. As persons who are interested in clinical grade cell manufacturing, we read the article, cited references and the authors’ previous publications to find out more details of their cell processing method. Our findings suggested that the cell manufacturing is STEM CELLS 2014;00:00‐00 www.StemCells.com
not fully cGMP compatible and needs to be improved. According to current regulations, cell‐based products should be manufactured under principles of cGMP when they are used in clinical studies. Besides cGMP facility (GMP condition), it is recommended to use FDA‐ approved or cleared, cGMP manufactured, or clinical grade reagents whenever they are available. If a re‐ search grade reagent is used as part of the manufactur‐ ing process, additional testing may be needed to ensure the safety and quality of the reagent [2]. Clinical grade cell manufacturing cannot be achieved by transfer of current methodology into a clean room [3]. The Authors ©AlphaMed Press 2014
2 applied collagenase type I for tissue digestion that seems to be research grade and its manufacturer is un‐ clear. Collagenase may contain mammalian‐based com‐ ponents that may carry infectious agents such as prions. Other types of collagenase such as: CLSAFA (Worthing‐ ton), NB 6 GMP Grade (SERVA), and Liberase MTF‐S GMP Grade (Roche), have been used for adipose tissue digestion. These clinical‐grade products can replace current research‐grade collagenase without any nega‐ tive effect in the yield or function of AD‐MSCs [4].The authors stated that FBS obtained from bovine spongi‐ form encephalopathy free herd. However, the manufac‐ turer of FBS, virus inactivation method (for example gamma irradiation) and its country of origin are unclear. If animal products cannot be replaced, we should use materials that are sourced in a controlled and docu‐ mented manner from animals bred and raised in captiv‐ ity in countries or geographic regions with appropriate national health status, disease prevention, and control systems [3]. The European Directorate for the Quality of Medicine & Healthcare (EDQM) has provided a list of TSE‐certified sera in its website [5]. We also could not find details of their passaging protocol. In clinical grade cell manufacturing, porcine trypsin should be replaced
REFERENCES 1. Jo CH, Lee YG, Shin WH, et al. Intra‐articular injection of mesenchymal stem cells for the treatment of osteoarthritis of the knee: A proof‐of‐concept clinical trial. Stem Cells.2014 ;doi: 10.1002/stem.1634. 2. Guidance for FDA Reviewers and Sponsors: Content and Review of Chemistry, Manufac‐ turing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs), Center for Biologics Evaluation and Research, April 2008. 3. Arjmand B, Emami‐Razavi SH, Larijani B, et al. The implementation of tissue banking
www.StemCells.com
by animal origin‐free alternatives such as: TrypLE™ Se‐ lect (Invitrogen) and TrypZean (Sigma‐aldrich). These reagents have been successfully used for passaging AD‐ MSCs and are gentle on cells [6].To propagate AD‐MSCs, rEGF supplemented keratinocyte‐SFM medium has been used in the study. Although rEGF and bFGF accel‐ erate AD‐MSC expansion and conserve the spindle shape morphology, it may limit its differentiation ability during ex vivo expansion. Therefore, these growth fac‐ tors should be carefully considered in stem cells for clinical applications [7]. Furthermore, Kølle et al., have described a simple, growth factor‐free GMP compatible protocol which yielded an average of 8.58 × 10⁸ AD‐ MSCs per 50 ml of fat tissue in 14 days [8]. We conclude that in cell based clinical trials besides considering good clinical practice (GCP), strict adherence to cGMP should be followed in cell manufacturing process.
AUTHOR CONTRIBUTIONS B.A. (literature review and data extraction) and H.R.A.: (manuscript preparation and correspondence)
experiences for setting up a cGMP cell manu‐ facturing facility. Cell Tissue Bank 2012; 13(4):587‐596. 4. Carvalho PP, Gimble JM, Dias IR, et al. Xeno free enzymatic products for the isolation of human adipose‐derived stromal/stem cells. Tissue Eng Part C Methods 2013;19(6):473‐ 478. 5. Reinhardt J, Stuhler A, Blumel J. Safety of Bovine Sera for Production of Mesenchymal Stem Cells for Therapeutic Use. Human Gene Therapy 2011; 22:775‐776. 6. Carvalho PP, Wu X, Yu G, Dietrich M, et al. Use of animal protein‐free products for pas‐ saging adherent human adipose‐derived
stromal/stem cells. Cytotherapy. 2011;13(5):594‐597. 7. Hu F, Wang X, Liang G, et al.Effects of epi‐ dermal growth factor and basic fibroblast growth factor on the proliferation and osteogenic and neural differentiation of adi‐ pose‐derived stem cells.Cell Reprogram 2013;15(3):224‐232. 8. Trojahn Kølle SF, Fischer‐Nielsen A, Mathiasen AB, et al. Enrichment of autolo‐ gous fat grafts with ex‐vivo expanded adipose tissue‐derived stem cells for graft survival: a randomised placebo‐controlled trial. The Lancet 2013; 382 (9898): 1113 – 1120.
©AlphaMed Press 2014
Cell Manufacturing for Clinical Applications BABAK ARJMAND1,2, HAMID REZA AGHAYAN2,3 Key words. AD‐MSCs • Cell manufacturing • GCP • GMP STEM CELLS 2014; 00:000–000
Corresponding author: Hamid Reza Aghayan, Endocrinology and Me‐ tabolism Research Center and GMP Cellular Therapy Facility, Chronic Diseases Research Center, Tehran University of Medical Sciences, Tehran, Iran. E‐Mail: [email protected] Received February 22, 2014; ac‐ cepted for publication March 16, 2014; available online without sub‐ scription through the open access option. ©AlphaMed Press 1066‐5099/2014/$30.00/0 This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typeset‐ ting, pagination and proofreading process which may lead to differ‐ ences between this version and the Version of Record. Please cite this article as doi: 10.1002/stem.1751 To the editor: The study of adipose tissue derived MSCs (AD‐MSCs) injection for osteoarthritis by Jo et al., was fascinating to us [1]. It is a promising study with good clinical trial design that contains valuable clinical information. In contrast to clinical data, cell manufacturing process has been briefly described. As persons who are interested in clinical grade cell manufacturing, we read the article, cited references and the authors’ previous publications to find out more details of their cell processing method. Our findings suggested that the cell manufacturing is STEM CELLS 2014;00:00‐00 www.StemCells.com
not fully cGMP compatible and needs to be improved. According to current regulations, cell‐based products should be manufactured under principles of cGMP when they are used in clinical studies. Besides cGMP facility (GMP condition), it is recommended to use FDA‐ approved or cleared, cGMP manufactured, or clinical grade reagents whenever they are available. If a re‐ search grade reagent is used as part of the manufactur‐ ing process, additional testing may be needed to ensure the safety and quality of the reagent [2]. Clinical grade cell manufacturing cannot be achieved by transfer of current methodology into a clean room [3]. The Authors ©AlphaMed Press 2014
2 applied collagenase type I for tissue digestion that seems to be research grade and its manufacturer is un‐ clear. Collagenase may contain mammalian‐based com‐ ponents that may carry infectious agents such as prions. Other types of collagenase such as: CLSAFA (Worthing‐ ton), NB 6 GMP Grade (SERVA), and Liberase MTF‐S GMP Grade (Roche), have been used for adipose tissue digestion. These clinical‐grade products can replace current research‐grade collagenase without any nega‐ tive effect in the yield or function of AD‐MSCs [4].The authors stated that FBS obtained from bovine spongi‐ form encephalopathy free herd. However, the manufac‐ turer of FBS, virus inactivation method (for example gamma irradiation) and its country of origin are unclear. If animal products cannot be replaced, we should use materials that are sourced in a controlled and docu‐ mented manner from animals bred and raised in captiv‐ ity in countries or geographic regions with appropriate national health status, disease prevention, and control systems [3]. The European Directorate for the Quality of Medicine & Healthcare (EDQM) has provided a list of TSE‐certified sera in its website [5]. We also could not find details of their passaging protocol. In clinical grade cell manufacturing, porcine trypsin should be replaced
REFERENCES 1. Jo CH, Lee YG, Shin WH, et al. Intra‐articular injection of mesenchymal stem cells for the treatment of osteoarthritis of the knee: A proof‐of‐concept clinical trial. Stem Cells.2014 ;doi: 10.1002/stem.1634. 2. Guidance for FDA Reviewers and Sponsors: Content and Review of Chemistry, Manufac‐ turing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs), Center for Biologics Evaluation and Research, April 2008. 3. Arjmand B, Emami‐Razavi SH, Larijani B, et al. The implementation of tissue banking
www.StemCells.com
by animal origin‐free alternatives such as: TrypLE™ Se‐ lect (Invitrogen) and TrypZean (Sigma‐aldrich). These reagents have been successfully used for passaging AD‐ MSCs and are gentle on cells [6].To propagate AD‐MSCs, rEGF supplemented keratinocyte‐SFM medium has been used in the study. Although rEGF and bFGF accel‐ erate AD‐MSC expansion and conserve the spindle shape morphology, it may limit its differentiation ability during ex vivo expansion. Therefore, these growth fac‐ tors should be carefully considered in stem cells for clinical applications [7]. Furthermore, Kølle et al., have described a simple, growth factor‐free GMP compatible protocol which yielded an average of 8.58 × 10⁸ AD‐ MSCs per 50 ml of fat tissue in 14 days [8]. We conclude that in cell based clinical trials besides considering good clinical practice (GCP), strict adherence to cGMP should be followed in cell manufacturing process.
AUTHOR CONTRIBUTIONS B.A. (literature review and data extraction) and H.R.A.: (manuscript preparation and correspondence)
experiences for setting up a cGMP cell manu‐ facturing facility. Cell Tissue Bank 2012; 13(4):587‐596. 4. Carvalho PP, Gimble JM, Dias IR, et al. Xeno free enzymatic products for the isolation of human adipose‐derived stromal/stem cells. Tissue Eng Part C Methods 2013;19(6):473‐ 478. 5. Reinhardt J, Stuhler A, Blumel J. Safety of Bovine Sera for Production of Mesenchymal Stem Cells for Therapeutic Use. Human Gene Therapy 2011; 22:775‐776. 6. Carvalho PP, Wu X, Yu G, Dietrich M, et al. Use of animal protein‐free products for pas‐ saging adherent human adipose‐derived
stromal/stem cells. Cytotherapy. 2011;13(5):594‐597. 7. Hu F, Wang X, Liang G, et al.Effects of epi‐ dermal growth factor and basic fibroblast growth factor on the proliferation and osteogenic and neural differentiation of adi‐ pose‐derived stem cells.Cell Reprogram 2013;15(3):224‐232. 8. Trojahn Kølle SF, Fischer‐Nielsen A, Mathiasen AB, et al. Enrichment of autolo‐ gous fat grafts with ex‐vivo expanded adipose tissue‐derived stem cells for graft survival: a randomised placebo‐controlled trial. The Lancet 2013; 382 (9898): 1113 – 1120.
©AlphaMed Press 2014
