Goldstein Ronald S
Faculty of Life Sciences, Bar-Ilan University, Gonda Research Center, Ramat-Gan, Israel.
Methods Mol Biol. 2010;584:367-85. doi: 10.1007/978-1-60761-369-5_20.
Traditional methods of studying the differentiation of human embryonic stem cells (hESCs) include generation of embryoid bodies, induced differentiation in vitro, and transplantation to immune-deficient mice. The chick embryo is a well-studied and accessible experimental system that has been used for many years as a xenograft host for mammalian cells. Several years ago, we performed experiments transplanting colonies of hESC into organogenesis-stage chick embryos to establish a novel system for studying the developmental programs and decisions of pluripotent human cells. Fluorescent hESC were used, in order to permit identification of the hESC in living embryos. We transplanted hESC into the trunk of chick embryos, both into and instead of developing somites. Our results showed that hESC survive, migrate, and integrate into the tissues of the chick embryo. Some of the hESC differentiated and the type of embryonic microenvironment that the implanted cells were exposed to modified their differentiation. Several other laboratories have subsequently xenografted hESC-derived cells to chick embryos for evaluating their differentiation in vivo. Therefore, the hESC-chick embryo system is a useful xenograft system complementing studies in rodents and in vitro, as well as uniquely shedding light on early processes in the development of human cells in the embryonic context.
研究人类胚胎干细胞(hESCs)分化的传统方法包括生成胚状体、体外诱导分化以及移植到免疫缺陷小鼠体内。鸡胚是一个经过充分研究且易于操作的实验系统,多年来一直被用作哺乳动物细胞的异种移植宿主。几年前,我们进行了将hESC集落移植到器官发生期鸡胚中的实验,以建立一个研究多能人类细胞发育程序和决定的新系统。使用了荧光hESC,以便在活胚胎中识别hESC。我们将hESC移植到鸡胚的躯干中,既移植到发育中的体节内,也移植到体节处替代体节。我们的结果表明,hESC能够存活、迁移并整合到鸡胚组织中。一些hESC发生了分化,植入细胞所接触的胚胎微环境类型改变了它们的分化。随后,其他几个实验室也将hESC衍生的细胞异种移植到鸡胚中,以评估它们在体内的分化情况。因此,hESC - 鸡胚系统是一个有用的异种移植系统,它补充了在啮齿动物和体外进行的研究,并且独特地揭示了胚胎环境中人类细胞发育的早期过程。