Efficient derivation of Chinese human embryonic stem cell lines from frozen embryos

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Date: Apr. 2010
Publisher: Springer
Document Type: Report
Length: 398 words

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Byline: Chunliang Li (1,5), Ying Yang (1,5), Xiaowei Lu (3), Yijuan Sun (3), Junjie Gu (1,2), Yun Feng (3), Ying Jin (1,2,4) Keywords: Human embryonic stem cells; Derivation; Characterization Abstract: Human embryonic stem (hES) cells are pluripotent cells derived from the inner cell mass of blastocysts. Their unique properties of self-renewal and pluripotency make them an attractive tool for basic research as well as a potential cell resource for therapy. However, each hES cell line demonstrates different identity. It is desirable to obtain more fully characterized hES cell lines with newly developed technologies associated with hES cell culture. Here, we report our experience of efficient derivation of three new Chinese hES cell lines (SHhES2, SHhES3, and SHhES4) from in vitro fertilization discarded embryos donated by women with polycystic ovary syndrome. These cell lines were derived under conditions minimizing exposure to animal components and maintained at an undifferentiated state for long-term culture. They retained a normal karyotype and expressed ALP, OCT4, SOX2, SSEA-4, TRA-1-60 and TRA-1-81. RT-PCR analysis also revealed high expression levels of pluripotency markers such as OCT4, LEFTY A, SOX2, TDGF-1, THY1, FGF4, NANOG, and REX1. When suspended in low-attachment culture dishes, embryoid bodies formed and were comprised of various differentiated cell types from all three embryonic germ layers. However, well-shaped teratomas were only harvested from line SHhES2, not from SHhES3 and SHhES4, indicating that the differentiation ability in vivo differs among the three cell lines. Collectively, the three new hES cell lines were established and fully characterized. The effort paves the way toward generating hES cell lines without contamination by animal components. All of these cell lines are available by contact Ying Jin at yjin@sibs.ac.cn. Author Affiliation: (1) Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, 225 South Chongqing Road, Shanghai, 200025, China (2) Shanghai Institute of Stem Cell Research, Shanghai Jiao Tong University School of Medicine, 225 South Chongqing Road, Shanghai, 200025, China (3) Reproductive Medical Center, Ruijin Hospital, 197 Second Ruijin Road, Shanghai, 200135, China (4) Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China (5) Graduate School of Chinese Academy of Sciences, Beijing, China Article History: Registration Date: 19/01/2010 Received Date: 26/03/2009 Accepted Date: 19/01/2010 Online Date: 26/02/2010 Article note: Editor: P. Andrews

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Gale Document Number: GALE|A231205616