期刊
NATURE PROTOCOLS
卷 8, 期 2, 页码 254-264出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2012.157
关键词
-
资金
- Flemish Foundation for Scientific Research (FWO- Vlaanderen) [FWO- 3G062910]
- Concerted Research Actions
- Bijzonder Onderzoeksfonds [BOF GOA 01G01112]
- Netherlands organization of Scientific Research (NWO) [ASPASIA 015.007.037]
- Interuniversity Attraction Poles (PAI) [P6/20]
Little is known about the true developmental origin of human embryonic stem cells (hESCs) or the events that initiate their generation. Recently, we have shown that hESCs originate from a post-inner cell mass (ICM) intermediate (PICMI), a unique transient epiblast-like structure that is different from both its ICM progenitor and its subsequent hESC fate. As a closer progenitor of hESCs than the ICM, the PICMI could be used to provide further insight into the human pluripotent state. Here we provide a detailed (7-d) protocol for the culture of the human preimplantation embryos in order to derive the PICMI. Subsequent identification and cryopreservation of the PICMI are described, in addition to hESC derivation. The initial hESC outgrowth is visible within 2-7 d after PICMI plating. By using the protocol provided, we observed PICMI formation in 21.3% of plated blastocysts with good-quality ICMs. Of the PICMIs used for hESC derivation, 80.6% showed hESC outgrowth after further culture.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据