Drissi Hicham, Gibson Jason D, Guzzo Rosa M, Xu Ren-He
Department of Orthopaedic Surgery, UConn Health, 263 Farmington Avenue, Farmington, CT, 06030-4037, USA.
Department of Genetics and Genome Sciences, UConn Health, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
Methods Mol Biol. 2015;1340:65-78. doi: 10.1007/978-1-4939-2938-2_5.
The induction of human embryonic stem cells to a mesenchymal-like progenitor population constitutes a developmentally relevant approach for efficient directed differentiation of human embryonic stem (hES) cells to the chondrogenic lineage. The initial enrichment of a hemangioblast intermediate has been shown to yield a replenishable population of highly purified progenitor cells that exhibit the typical mesenchymal stem cell (MSC) surface markers as well as the capacity for multilineage differentiation to bone, fat, and cartilage. Herein, we provide detailed methodologies for the derivation and characterization of potent mesenchymal-like progenitors from hES cells and describe in vitro assays for bone morphogenetic protein (BMP)-2-mediated differentiation to the chondrogenic lineage.
将人类胚胎干细胞诱导为间充质样祖细胞群体,是一种将人类胚胎干(hES)细胞高效定向分化为软骨生成谱系的与发育相关的方法。已证明,最初富集成血管细胞中间体可产生可补充的高度纯化祖细胞群体,这些祖细胞表现出典型的间充质干细胞(MSC)表面标志物,以及向骨、脂肪和软骨多谱系分化的能力。在此,我们提供了从hES细胞中获得高效间充质样祖细胞及其特性鉴定的详细方法,并描述了骨形态发生蛋白(BMP)-2介导的向软骨生成谱系分化的体外测定方法。