McElroy Sohyun L, Reijo Pera Renee A
Center for Human Embryonic Stem Cell Research and Education, Institute for Stem Cell Biology and Regenerative Medicine, Department of Obstetrics and Gynecology, Stanford University, Palo Alto, CA 94304-5542, USA.
CSH Protoc. 2008 Sep 1;2008:pdb.prot5041. doi: 10.1101/pdb.prot5041.
INTRODUCTIONEmbryonic stem cells (ESCs) are derived from the inner cell mass of day 5-6 blastocysts. ESCs are pluripotent, meaning that they are able to differentiate into all derivatives of the three primary germ layers (ectoderm, endoderm, and mesoderm). In order to maintain the undifferentiated status of human ESCs (hESCs), feeder cells are used to provide both a suitable attachment substrate and critical soluble factors. Since the first hESC lines were established on mouse embryonic fibroblasts (MEFs), mitotically inactivated MEFs have commonly been used for supporting the culture of undifferentiated hESCs. Some previous studies suggest that MEFs may support hESC growth better than the human feeder cells typically isolated from post-natal tissues. This protocol describes a method for isolation and irradiation of MEFs for use in hESC culture.
引言
胚胎干细胞（ESC）来源于第5 - 6天囊胚的内细胞团。胚胎干细胞具有多能性，这意味着它们能够分化为三个原始胚层（外胚层、内胚层和中胚层）的所有衍生物。为了维持人类胚胎干细胞（hESC）的未分化状态，饲养层细胞用于提供合适的附着底物和关键的可溶性因子。自从第一批hESC系在小鼠胚胎成纤维细胞（MEF）上建立以来，有丝分裂失活的MEF通常被用于支持未分化hESC的培养。一些先前的研究表明，MEF可能比通常从产后组织中分离的人类饲养层细胞更能支持hESC的生长。本方案描述了一种用于hESC培养的MEF分离和辐照方法。
