NWIPB OpenIR
Establishment of a feeder and serum-free culture system for human embryonic stem cells
Wang, LiYun; Zhang, RuiNa; Ma, RongHua; Jia, GongXue; Jian, ShengYan; Zeng, XiangHui; Xiong, ZhengFang; Li, BinYe; Li, Chen; Lv, ZhenZhen; Bai, Xue
2020
Source PublicationZYGOTE
Volume28Issue:3
AbstractStem cells are an immortal cell population capable of self-renewal; they are essential for human development and ageing and are a major focus of research in regenerative medicine. Despite considerable progress in differentiation of stem cellsin vitro, culture conditions require further optimization to maximize the potential for multicellular differentiation during expansion. The aim of this study was to develop a feeder-free, serum-free culture method for human embryonic stem cells (hESCs), to establish optimal conditions for hESC proliferation, and to determine the biological characteristics of the resulting hESCs. The H9 hESC line was cultured using a homemade serum-free, feeder-free culture system, and growth was observed. The expression of pluripotency proteins (OCT4, NANOG, SOX2, LIN28, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81) in hESCs was determined by immunofluorescence and western blotting. The mRNA expression levels of genes encoding nestin, brachyury and alpha-fetoprotein in differentiated H9 cells were determined by RT-PCR. The newly developed culture system resulted in classical hESC colonies that were round or elliptical in shape, with clear and neat boundaries. The expression of pluripotency proteins was increased, and the genes encoding nestin, brachyury, and alpha-fetoprotein were expressed in H9 cells, suggesting that the cells maintainedin vitrodifferentiation capacity. Our culture system containing a unique set of components, with animal-derived substances, maintained the self-renewal potential and pluripotency of H9 cells for eight passages. Further optimization of this system may expand the clinical application of hESCs.
KeywordBasic fibroblast growth factor Feeder-free Human embryonic stem cells Pluripotency Self-renewal
Document Type期刊论文
Identifierhttp://210.75.249.4/handle/363003/60320
Collection中国科学院西北高原生物研究所
Recommended Citation
GB/T 7714
Wang, LiYun,Zhang, RuiNa,Ma, RongHua,et al. Establishment of a feeder and serum-free culture system for human embryonic stem cells[J]. ZYGOTE,2020,28(3).
APA Wang, LiYun.,Zhang, RuiNa.,Ma, RongHua.,Jia, GongXue.,Jian, ShengYan.,...&Bai, Xue.(2020).Establishment of a feeder and serum-free culture system for human embryonic stem cells.ZYGOTE,28(3).
MLA Wang, LiYun,et al."Establishment of a feeder and serum-free culture system for human embryonic stem cells".ZYGOTE 28.3(2020).
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