Feeder-free growth of undifferentiated human embryonic stem cells

被引：1378

作者：

Xu, CH ^{[1
]}

Inokuma, MS ^{[1
]}

Denham, J ^{[1
]}

Golds, K ^{[1
]}

Kundu, P ^{[1
]}

Gold, JD ^{[1
]}

Carpenter, MK ^{[1
]}

机构：

[1] Geron Corp, Menlo Pk, CA 94025 USA

来源：

NATURE BIOTECHNOLOGY | 2001年 / 19卷 / 10期

关键词：

D O I：

10.1038/nbt1001-971

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Previous studies have shown that maintenance of undifferentiated human embryonic stem (hES) cells requires culture on mouse embryonic fibroblast (MEF) feeders. Here we demonstrate a successful feeder-free hES culture system in which undifferentiated cells can be maintained for at least 130 population doublings. In this system, hES cells are cultured on Matrigel or laminin in medium conditioned by MEF. The hES cells maintained on feeders or off feeders express integrin alpha6 and beta1, which may form a laminin-specific receptor. The hES cell populations in feeder-free conditions maintained a normal karyotype, stable proliferation rate, and high telomerase activity. Similar to cells cultured on feeders, hES cells maintained under feeder-free conditions expressed OCT-4, hTERT alkaline phosphatase, and surface markers including SSEA-4, Tra 1-60, and Tra 1-81. In addition, hES cells maintained without direct feeder contact formed teratomas in SCID/beige mice and differentiated in vitro into cells from all three germ layers. Thus, the cells retain fundamental characteristics of hES cells in this culture system and are suitable for scaleup production.

引用

页码：971 / 974

页数：4

共 15 条

[1] Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture [J].