Lin Ge, Martins-Taylor Kristen, Xu Ren-He
Department of Genetics and Developmental Biology, Stem Cell Institute, University of Connecticut Health Center, Farmington, CT, USA.
Methods Mol Biol. 2010;636:1-24. doi: 10.1007/978-1-60761-691-7_1.
Since the first report of derivation of human embryonic stem cell (hESC) lines in 1998, many progresses have been achieved to reliably and efficiently derive, maintain, and differentiate this therapeutically promising cell type. This chapter introduces some basic and widely recognized methods that we use in our hESC core laboratory. Specifically, it includes methods for (1) deriving hESC lines without using enzyme and antibody to isolate the inner cell mass; (2) sustaining hESC self-renewal under feeder-dependent, feeder-conditioned, and defined conditions as well as pluripotency validation and quality control assays; and (3) inducing hESC differentiation to trophoblast with BMP4.
自1998年首次报道人类胚胎干细胞(hESC)系的衍生以来,在可靠且高效地衍生、维持和分化这种具有治疗前景的细胞类型方面已取得了许多进展。本章介绍了我们在hESC核心实验室中使用的一些基本且被广泛认可的方法。具体而言,它包括以下方法:(1)在不使用酶和抗体分离内细胞团的情况下衍生hESC系;(2)在依赖饲养层、饲养层条件培养基和限定条件下维持hESC自我更新以及多能性验证和质量控制检测;(3)用BMP4诱导hESC分化为滋养层细胞。