Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
Diagnostic/Biological Sciences, School of Dentistry, University of Minnesota, Minneapolis, MN 55455, USA.
Stem Cell Reports. 2015 Feb 10;4(2):190-8. doi: 10.1016/j.stemcr.2015.01.008.
We generated a RUNX2-yellow fluorescent protein (YFP) reporter system to study osteogenic development from human embryonic stem cells (hESCs). Our studies demonstrate the fidelity of YFP expression with expression of RUNX2 and other osteogenic genes in hESC-derived osteoprogenitor cells, as well as the osteogenic specificity of YFP signal. In vitro studies confirm that the hESC-derived YFP(+) cells have similar osteogenic phenotypes to osteoprogenitor cells generated from bone-marrow mesenchymal stem cells. In vivo studies demonstrate the hESC-derived YFP(+) cells can repair a calvarial defect in immunodeficient mice. Using the engineered hESCs, we monitored the osteogenic development and explored the roles of osteogenic supplements BMP2 and FGF9 in osteogenic differentiation of these hESCs in vitro. Taken together, this reporter system provides a novel system to monitor the osteogenic differentiation of hESCs and becomes useful to identify soluble agents and cell signaling pathways that mediate early stages of human bone development.
我们构建了 RUNX2-黄色荧光蛋白（YFP）报告系统，用于研究人胚胎干细胞（hESC）向成骨细胞的分化。我们的研究表明，YFP 表达与 hESC 来源的成骨前体细胞中 RUNX2 和其他成骨基因的表达具有很好的相关性，且 YFP 信号具有成骨细胞特异性。体外研究证实，hESC 来源的 YFP(+)细胞具有与骨髓间充质干细胞来源的成骨前体细胞相似的成骨表型。体内研究表明，hESC 来源的 YFP(+)细胞可修复免疫缺陷小鼠的颅骨缺损。利用工程化的 hESC，我们监测了成骨细胞的分化，并探索了骨形态发生蛋白 2（BMP2）和成纤维细胞生长因子 9（FGF9）等成骨补充剂在体外诱导 hESC 成骨分化中的作用。综上所述，该报告系统为监测 hESC 的成骨分化提供了一种新方法，有助于鉴定可介导人骨早期发育的可溶性因子和细胞信号通路。
