Kramer Jan, Hargus Gunnar, Rohwedel Jürgen
Department of Medical Molecular Cell Biology, University of Lübeck, Germany.
Methods Mol Biol. 2006;330:171-90. doi: 10.1385/1-59745-036-7:171.
The model system of embryonic stem (ES) cell differentiation in vitro via cellular aggregates (embryoid bodies, EBs) can be used to analyze cell differentiation from a pluripotent stem cell via progenitor cells up to terminally differentiated cell types. ES cells are known to be pluripotent; they have the capacity to differentiate into any cell lineage of the three germ layers. Using various ES cell lines, we characterized chondrogenic and osteogenic differentiation in EBs by histochemical staining, immunostaining, mRNA-in situ hybridization, and reverse transcriptase polymerase chain reaction analysis. Here, we describe in detail our established protocols to analyze chondrogenic differentiation of ES cells. We summarize different ways to modulate ES cell-derived chondrogenic differentiation.
通过细胞聚集体(胚状体,EBs)进行胚胎干细胞(ES细胞)体外分化的模型系统,可用于分析从多能干细胞经祖细胞直至终末分化细胞类型的细胞分化过程。已知ES细胞具有多能性;它们有能力分化为三个胚层的任何细胞谱系。我们使用各种ES细胞系,通过组织化学染色、免疫染色、mRNA原位杂交和逆转录聚合酶链反应分析,对EBs中的软骨生成和成骨分化进行了表征。在此,我们详细描述了我们建立的用于分析ES细胞软骨生成分化的方案。我们总结了调节ES细胞来源的软骨生成分化的不同方法。