Figure 2.

Colony-forming unit fibroblasts for ESC_endo and ESC_cyst after they were cultured at clonal density for 21 days. Also, osteoblast differentiation for ESC_endo and ESC_cyst following 14–21 days culture in osteoblast differentiation growth medium and adipocyte differentiation for ESC_endo and ESC_cyst following 28-day culture in adipocyte differentiation growth medium are shown. The colony-forming efficiency (a) shows that ESC_endo formed significantly more colonies than the ESC_cyst (^∗P < 0.05). Representative phase contrast images (b) at 4x magnification following Eosin staining of ESC_endo and ESC_cyst. Colonies with ≥50 cells were counted. Quantitation of the Alizarin red S dye (c) as a mean fold change relative to the untreated controls; both types of stromal cells significantly induced osteoblast differentiation (^∗P < 0.05). Representative phase contrast images (d) at 20x magnification following Alizarin red S staining of the calcium salts. Quantitation of the Oil red O dye (e) as a mean fold change relative to the untreated controls; both types of stromal cells significantly induced adipocyte differentiation (^∗P < 0.05). Representative phase contrast images (f) at 20x magnification following Oil red O staining of the lipid vacuoles. Nine independent experiments (n = 3 biological replicates) were carried out in triplicates for (a, b). Three to four independent experiments (n = 3 biological replicates) were carried out in duplicates for (c, d, e, f). Mean ± SD. Scale bars represent 50 μm at 4x magnification for (a,b) and 50 μm at 20x magnification for (c, d, e, f).