Tri-lineage differentiation potential of tissue cells. Cells from bone marrow (BM) (A-C), breast adipose (BA) (D-F), foreskin fibroblasts (FF) (G-I) and olfactory tissue (OT) (J-L), and were exposed to conditions known to induce the differentiation of mesenchymal stem cells into chondrocytes, adipocytes and osteocytes. Cytochemical stains were carried out to indicate differentiation; Safranin O (chondrocytes: A, D, G, J-Scale bar (J) = 200 μm), Oil-red-O (adipocytes: B, E, H, K whole well scans (5×5 montage) Scale bar (K) = 1 mm: inset-Scale bar (K) = 40 μm) and Alizarin Red (osteocytes: C, F-Scale bar (F) = 100 μm: I, L-Scale bar (L) = 50 μm). All images are representative except for F and L which portray very rare osteocyte staining in FF and OT cultures. M: Illustrates confirmation and quantitation of differentiation by qPCR. Taqman probes for aggrecan, adiponectin and osteopontin were used to quantitate chondrocyte, adipocyte and osteocyte differentiation respectively amongst our tissue samples. ΔΔCt values were calculated relative to GAPDH and were normalized relative to BM (N = 3/cell type). Standard error is displayed and significant differences between tissue types (P < 0.05) are indicated (*). RNA was extracted on the same day as cytochemical analysis; chondrocytes (day 21), adipocytes (day 25) and osteocytes (day 21).
Wetzig et al. BMC Cell Biology 2013 14:54 doi:10.1186/1471-2121-14-54