High throughput screening to identify Runx2 independent osteogenesis

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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.