Adipogenic and osteogenic differentiation.

<p>(A) Total RNA was isolated from three MSC samples cultured on TC surface (CON) or COL, FBN or PLL coated surfaces and reverse transcribed. The mRNA expression levels of OCN and PPARG were analysed by real-time PCR and normalized to their respective GAPDH levels. Values are Mean±SE, n = 3 samples. MSC were isolated on uncoated TC surface and differentiated into either (B, C) adipocytes or (D, E) osteocytes on different matrices (Control, COL, FBN and PLL) for 21–35 days. (B, C) To detect adipogenic differentiation, the cells were stained with oil-red O and the positive cells were counted microscopically. (D, E) Osteogenic differentiation was determined by staining the cells with Alizarin Red and quantified by absorbance measurement at 562nm. Values are Mean±SD, n = 6–12. (F) Osteogenic differentiation during switching of cell culture surface. MSC were cultured for one week prior to induction of osteogenic differentiation on control TC treated surface (CON), COL, FBN or PLL. These cells were transferred to and differentiated into osteocytes either on COL, FBN or PLL surfaces and osteogenic differentiation was determined alizarin red staining. For example, COL-FBN means that the cells were cultured on COL and switched to FBN for osteogenic differentiation. Values are Mean+SD, n = 3 independent experiments.*p<0.05, **p<0.005, ***p<0.0005.