Fig. 4.

Separation and identification of BMMSCs and exosomes. A Morphological characteristics of BMMSCs (× 100); (a) primary cells; (b) cells after 3-day culture; (c) cells after 12-day culture; (d) P3 generation cells. B Surface markers of BMMSCs determined by flow cytometry. C Osteogenesis- and adipogenesis-inducing cultures of BMMSCs; (a) osteogenic differentiation, (b) adipogenic differentiation (× 400). D Exosomes identified by transmission electron microscope (× 50,000). E Diameter distribution of BMMSC-derived exosomes measured by Image-Pro plus software. F Expression of exosomal surface marker CD63 determined by flow cytometry. G Levels of CD63, TSG101, and calnexin examined by western blot analysis. *p < 0.05 versus cell lysis buffer. The measurement data were described as mean ± standard deviation. Independent-sample t-test was employed to analyze difference between two groups. The cell experiment was repeated three times