High throughput screening to identify Runx2 independent osteogenesis

携带Runx2基因缺陷的小鼠具有胚胎致死性，这种基因突变导致人类锁骨颅发育不良（CCD）。因此，已知Runx2对成骨细胞分化很重要。目前，我们正在利用人类和小鼠iPSCs研究不依赖runx2的成骨作用。我们已经分析了几种可能在没有Runx2基因的情况下也能诱导成骨的试剂。在不同浓度维甲酸的实验中，我们证实了runx2阴性细胞中成骨基因的表达。由于成骨分化遵循时间差异表达，我们还分析了不同时间点的成骨基因表达，以确认我们当前分析的准确性。我们目前正在使用我们的研究方案进行钙化和矿化试验来评估骨形成能力。

The mouse with Runx2 gene defect is embryonically lethal, and this gene mutation causes cleidocranial dysplasia (CCD) in humans. Thus, Runx2 isknown to be important for osteoblast differentiation.We are currently investigating Runx2-independent osteogenesis using human and mouse iPSCs. We have analyzed several potential reagents that caninduce osteogenesis even in the absence of the Runx2 gene. In our experiment with retinoic acid at different concentrations, we confirmed theexpression of osteogenic genes in Runx2-negative cells. Because osteogenic differentiation follows differential temporal expression, we alsoanalyzed osteogenic gene expression at different time points to confirm the accuracy of our current analysis. We are currently performing thecalcification and mineralization assay to assess bone formation ability using our study protocol.