Preosteoblasts Enhance the Proliferation and Osteogenesis of Embryonic Stem Cells via Cell—Cell Contact and Osteogenic Supplements

摘要

Embryonic stem cells (ESCs) have been considered to be a promising cell type for regenerative medicine and tissue engineering applications, by virtue of their great capacity for self-renewal and potential for differentiation into cells of various types of tissues. However, the role of osteoblasts during ESC differentiating into the osteogenesis is still unknown. In this study, a coculture model was developed by using C3H10T1/2, a murine embryonic cell line, and MC3T3-E1, a murine preosteoblast cell line. These two cells were well mixed and cultured based on different ratios. To observe and compare the effect of osteogenic supplements on the proliferation and maturation of C3H10T1/2 cells during the osteogenic differentiation, dexamethason, L-ascorbic acid, and p-glycerophosphate were additional added in to the culture medium. Our results showed that cells cocultured in the osteogenic medium expressed higher proliferation than in the basal medium. Except C5 group (C3H10T1/2 cells only), all the mixed groups showed similar total alkaline phosphatase (ALP) activity in the osteogenic and basal medium. The total ALP activity of C5 group was significantly higher under the osteogenic condition. Coculture groups with higher ratios of MC3T3-E1 cells appeared more significant ALP activity and calcium deposit by ALP staining and alizarin red S staining, respectively during the coculture period.