马胚胎干细胞的体外衍生、维持及分化诱导

Derivation, maintenance, and induction of the differentiation in vitro of equine embryonic stem cells.

作者信息

Saito Shigeo, Sawai Ken, Minamihashi Arika, Ugai Hideyo, Murata Takehide, Yokoyama Kazunari K

机构信息

Saito Laboratory of Cell Technology, Kataoka, Yaita, Techigi, Japan.

出版信息

Methods Mol Biol. 2006;329:59-79. doi: 10.1385/1-59745-037-5:59.

DOI:10.1385/1-59745-037-5:59

PMID:16845984

Abstract

We describe here the isolation and maintenance of pluripotent embryonic stem (ES) cells from equine blastocysts that have been frozen and thawed. Equine ES cells appear to maintain a normal diploid karyotype in culture. These cells express markers that are characteristic of mouse ES cells, namely, alkaline phosphatase, stage-specific-embryonic antigen 1, STAT3, and Oct4. We also describe protocols for the induction of differentiation in vitro to neural precursor cells in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor, and platelet-derived growth factor and to hematopoietic and endothelial cell lineages in the presence of bFGF, stem cell factor, and oncostatin M. Equine ES cells provide a powerful tool for gene targeting and the generation of transgenic clonal offspring.

摘要

我们在此描述了从经冷冻和解冻的马囊胚中分离和培养多能胚胎干细胞（ES细胞）的方法。马ES细胞在培养中似乎保持正常的二倍体核型。这些细胞表达小鼠ES细胞特有的标志物，即碱性磷酸酶、阶段特异性胚胎抗原1、信号转导和转录激活因子3（STAT3）以及八聚体结合转录因子4（Oct4）。我们还描述了在碱性成纤维细胞生长因子（bFGF）、表皮生长因子和血小板衍生生长因子存在下体外诱导分化为神经前体细胞，以及在bFGF、干细胞因子和制瘤素M存在下诱导分化为造血和内皮细胞谱系的方案。马ES细胞为基因靶向和转基因克隆后代的产生提供了一个强大的工具。