Abstract
The purpose of this study is to evaluate osteogenic differentiation of dedifferentiated fat cells (DFATs) in 3D culture with RADA16, a self-assembling peptide. DFATs derived from rabbit subcutaneous fat were cultured in RADA16 in either a control medium (CM) or an osteogenic medium (OM) containing dexamethasone, L-ascorbic acid 2-phosphate, and β-glycerophosphate for 21 days. The cells cultured in the OM showed significantly higher DNA content on days 7 and 21, significantly higher osteocalcin content on days 7 and 14, and significantly higher calcium content on days 7, 14, and 21, compared to the cells cultured in CM. Von Kossa staining revealed the formation of a mineralized extracellular matrix with calcium deposition on days 14 and 21 of culture. These findings demonstrate that DFATs cultured in RADA16 differentiate into osteoblasts as early as day 7 of culture to form cultured bone in vitro. RADA16 promotes proliferation and differentiation of DFATs and thus might be a useful scaffold in bone tissue engineering.
