Behboodi Esmail, Lam Louise, Gavin William G, Bondareva Alla, Dobrinski Ina
Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2013;1074:51-67. doi: 10.1007/978-1-62703-628-3_5.
Embryonic stem (ES) cells are derived from the inner cell masses of preimplantation embryos. ES cells are pluripotent cells with the capacity for long-term propagation and broad differentiation plasticity. These cells have an exceptional functional feature in that they can differentiate into all tissues and organs, including germ cells. Established ES cell lines have been generated in mouse, human, and nonhuman primate but derivation of ES cells in farm animals has been problematic. Several ES-like cell lines from farm animals have been reported to exhibit properties of pluripotency in vitro. However, only a few of them morphologically resemble ES cells, or express markers that are associated with established ES cell lines from mouse and humans. Methods for derivation, propagation, and differentiation of ES cells from domestic animals have not been fully established. In this chapter, we describe methods for isolation of goat ES (gES) cell lines from in vivo-derived blastocysts and characterization of markers indicative of pluripotency. In addition, we outline differentiation of gES cells into all three germ layers in vivo by forming teratomas as a hallmark of pluripotency.
胚胎干细胞（ES细胞）来源于植入前胚胎的内细胞团。ES细胞是具有长期增殖能力和广泛分化可塑性的多能细胞。这些细胞具有特殊的功能特性，即它们可以分化为包括生殖细胞在内的所有组织和器官。已在小鼠、人类和非人类灵长类动物中建立了ES细胞系，但在农场动物中ES细胞的衍生一直存在问题。据报道，几种来自农场动物的类ES细胞系在体外表现出多能性。然而，其中只有少数在形态上类似于ES细胞，或表达与已建立的小鼠和人类ES细胞系相关的标志物。从家畜中衍生、增殖和分化ES细胞的方法尚未完全建立。在本章中，我们描述了从体内来源的囊胚中分离山羊ES（gES）细胞系以及表征多能性指示标志物的方法。此外，我们概述了通过形成畸胎瘤作为多能性标志，将gES细胞在体内分化为所有三个胚层的过程。
