Figure 2

Cells-directedin vitrodifferentiation of the CAF and the NTF. A-B. Adipogenic differentiation. Nile Red staining of lipid deposition (arrow indicated) in the cytoplasm (A). RTqPCR result showed the mean of PPARG gene regulation (B). C-D. Osteogenic differentiation. Alkaline phosphatase staining of macro- (upper panel) and microscopic visualization (lower panel). Scale bar, 100 μm (C). RTqPCR result of osteogenic differentiation genes osteocalcin (BGLAP), bone sialoprotein (IBSP), osteopontin (OPN), osteoprotegerin (OPG) and osteonectin (ON) (D). E-G. Endodermic differentiation. The qualitative result of the induction of somatostation (SST) and gastric inhibitory protein (GIP) using gel electrophoresis (E). RTqPCR result of endodermic differentiation genes showed an induction/up-regulation of SST and GIP. NIC = nicotinamide. The target mRNA expression was normalized to reference gene β-actin (F). Confirmation by DNA sequencing of human SST. SST_HCC3 = DNA sequences of SST from induced CAF, NM_001048.3 = reference DNA sequences from GenBank (G). For RTqPCR data, basal expression was considered as 1.00, target mRNA expressions were normalized to reference genes RNA18S and ACTB. Scale bar, 100 μm. *p < 0.05.