Jackson Melany, Taylor A Helen, Jones Elizabeth A, Forrester Lesley M
John Hughes Bennett Laboratory, Queen's Medical Research Institute, University of Edinburgh, EH164TJ, Edinburgh, UK.
Methods Mol Biol. 2010;633:1-18. doi: 10.1007/978-1-59745-019-5_1.
Embryonic stem (ES) cells are pluripotent cells isolated from the inner cell mass of the pre-implantation blastocyst. They have the capacity to undergo indefinite rounds of self-renewing cell division and differentiate into all the cell lineages of the developing embryo. In suspension culture, ES cells will differentiate into aggregates known as embryoid bodies in a manner similar to the early embryo. This culture system therefore provides a useful model to study the relatively inaccessible stages of mammalian development. We describe methods for the routine maintenance of mouse embryonic stem cells in culture, assays of stem cell self-renewal potential in monolayer culture and the generation of embryoid bodies to study differentiation pathways.
胚胎干细胞(ES细胞)是从植入前囊胚的内细胞团中分离出来的多能细胞。它们能够进行无限轮次的自我更新细胞分裂,并分化为发育中胚胎的所有细胞谱系。在悬浮培养中,ES细胞会以类似于早期胚胎的方式分化为称为类胚体的聚集体。因此,这种培养系统为研究哺乳动物发育中相对难以接近的阶段提供了一个有用的模型。我们描述了在培养中常规维持小鼠胚胎干细胞的方法、单层培养中干细胞自我更新潜力的测定以及生成类胚体以研究分化途径的方法。