Fernandes Karl J L, Miller Freda D
Département de pathologie et biologie cellulaire, Université de Montréal, Montréal, Québec, Canada.
Methods Mol Biol. 2009;482:159-70. doi: 10.1007/978-1-59745-060-7_10.
The isolation and experimental manipulation of multipotent precursors is of increasing therapeutic relevance. We recently reported the generation of cultures of Skin-derived Precursors ('SKPs'), multipotent cells that can be isolated from the dermis of embryonic, neonatal, and adult rodent skin (1), and from adult human skin (2) SKPs have similarities to stem cells of the embryonic neural crest (3), and differentiate into a variety of neural and mesodermal cell phenotypes, including peripheral neurons and glial cells, smooth muscle cells, bone, cartilage, and adipocytes (3-5). Here, we detail the establishment, propagation, neural differentiation, and immunocytochemical analysis of SKP cultures.
多能前体细胞的分离和实验操作具有越来越重要的治疗意义。我们最近报道了皮肤衍生前体细胞(“SKP”)培养物的生成,SKP是一种多能细胞,可以从胚胎、新生和成年啮齿动物皮肤的真皮中分离出来(1),也可从成人皮肤中分离出来(2)。SKP与胚胎神经嵴干细胞有相似之处(3),并能分化为多种神经和中胚层细胞表型,包括外周神经元和神经胶质细胞、平滑肌细胞、骨、软骨和脂肪细胞(3 - 5)。在此,我们详细介绍SKP培养物的建立、增殖、神经分化及免疫细胞化学分析。