title: Generation of clinical-grade human induced pluripotent stem cells in Xeno-free conditions.
abstract: BACKGROUND
  Human induced pluripotent stem cells (hiPSCs) are considered as one of the most promising seed cell sources in regenerative medicine. Now hiPSC-based clinical trials are underway. To ensure clinical safety, cells used in clinical trials or therapies should be generated under GMP conditions, and with Xeno-free culture media to avoid possible side effects like immune rejection that induced by the Xeno reagents. However, up to now there are no reports for hiPSC lines developed completely under GMP conditions using Xeno-free reagents.
  
  
  METHODS
  Clinical-grade human foreskin fibroblast (HFF) cells used as feeder cells and parental cells of the clinical-grade hiPSCs were isolated from human foreskin tissues and cultured in Xeno-free media. Clinical-grade hiPSCs were derived by integration-free Sendai virus-based reprogramming kit in Xeno-free pluriton™ reprogramming medium or X medium. Neural cells and cardiomyocytes differentiation were conducted following a series of spatial and temporal specific signals induction according to the corresponding lineage development signals. Biological safety evaluation of the clinical-grade HFF cells and hiPSCs were conducted following the guidance of the "Pharmacopoeia of the People's Republic of China, Edition 2010, Volume III".
  
  
  RESULTS
  We have successfully derived several integration-free clinical-grade hiPSC lines under GMP-controlled conditions and with Xeno-free reagents culture media in line with the current guidance of international and national evaluation criteria. As for the source of hiPSCs and feeder cells, biological safety evaluation of the HFF cells have been strictly reviewed by the National Institutes for Food and Drug Control (NIFDC). The hiPSC lines are pluripotent and have passed the safety evaluation. Moreover, one of the randomly selected hiPSC lines was capable of differentiating into functional neural cells and cardiomyocytes in Xeno-free culture media.
  
  
  CONCLUSIONS
  The clinical-grade hiPSC lines therefore could be valuable sources for future hiPSC-based clinical trials or therapies and for drug screening.
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. juan920846643@163.com.|College of Life Science, Northeast Agricultural University of China, Harbin, 150030, China. juan920846643@163.com.
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. haojie@ioz.ac.cn.
authors.forename: Jie
authors.initials: J
authors.lastname: Hao
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. donghuibai1989@163.com.
authors.forename: Donghui
authors.initials: D
authors.lastname: Bai
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. qizhi502@163.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Gu
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. 675657857@qq.com.|Graduate School of Chinese Academy of Sciences, Beijing, 100049, China. 675657857@qq.com.
authors.forename: Weifang
authors.initials: W
authors.lastname: Han
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. 191446410@qq.com.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. tanyuanq@sohu.com.|Graduate School of Chinese Academy of Sciences, Beijing, 100049, China. tanyuanq@sohu.com.
authors.forename: Yuanqing
authors.initials: Y
authors.lastname: Tan
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. 1214079962@qq.com.|College of Life Science, Northeast Agricultural University of China, Harbin, 150030, China. 1214079962@qq.com.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. xueke777@126.com.
authors.forename: Ke
authors.initials: K
authors.lastname: Xue
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. hpcsoft88@aliyun.com.
authors.forename: Pencheng
authors.initials: P
authors.lastname: Han
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. 599141004@qq.com.
authors.forename: Zhengxin
authors.initials: Z
authors.lastname: Liu
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. jiayd@ioz.ac.cn.
authors.forename: Yundan
authors.initials: Y
authors.lastname: Jia
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. wuxf@ioz.ac.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. liul@ioz.ac.cn.
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. wangliu@ioz.ac.cn.
authors.forename: Liu
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. liwei@ioz.ac.cn.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: College of Life Science, Northeast Agricultural University of China, Harbin, 150030, China. liu086@163.com.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
authors.affiliation: State Key of Stem Cells and Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. zhouqi@ioz.ac.cn.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D065150:Cellular Reprogramming; D003470:Culture Media / Q000737:chemistry*; D061252:Feeder Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007202:Indicators and Reagents; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D008297:Male; D032383:Myocytes, Cardiac / Q000166:cytology; D009474:Neurons / Q000166:cytology; D044968:Regenerative Medicine; D012449:Safety
chemical_list: D003470:Culture Media; D007202:Indicators and Reagents