In vitro osteogenic, adipogenic differentiation and gene expression of iPSC-cells generated using different factors. (A) Osteogenic differentiation of iPSC-cells-derived by exposure to RA alone; bFGF-RA, TGF-beta1-RA and TGF-beta3-RA. (B) quantitation of Alizarin Red deposition by each of the cell population. iPSC-TGF-beta1-RA-derived cells deposited more calcium based on Alizarin Red quantitation. (C) Expression of osteoblasts associated genes by differentiating cell populations. iPSC-BMP-2-RA cells express lower levels of osteogenic genes; (D) Adipocytic differentiation of iPSC- RA-cells, bFGF-RA-cells, TGF-beta1,3-RA-cells and BMP-2-RA-cells as assessed by Oil Red O staining. (D) All cells show adipogenic differentiation but BMP-2 derived cells show higher adipogenesis. (E) Quantitation of adipogenic differentiation based on Oil Red O deposition; all cells show adipogenic differentiation but BMP-2 derived cells show higher adipogenesis. (F) Semi quantitative PCR of osteoblasts and adipocytes associated genes expression by the iPSC-cells respectively. BMP-2-derived iPSC-cells showed least expression of osteoblasts associated genes but higher adipogenic differentiation markers. P <0.05. NM= normal medium, OM= osteogenic medium, AM= adipogenic medium. OSX= Osterix, OPN= osteopontin, OCN= Osteocalcin.