Abstract: Objective To study the proliferation characteristics of murine muscle-derived stem cells (MDSCs) in three different media. Methods MDSCs were isolated using mixed enzyme digestion combined with magnetic-activated cell sorting. Cells were divided into three groups, which were respectively cultured in normal media (NM), chicken embryo extract-containing media (CEECM) and cytokine-containing media (CKCM). The morphology of the cells was observed by phase contrast microscopy. The proliferation activity of the cells was detected by CCK8. The expression of proliferating cell nuclear antigen (PCNA), stem cell antigen 1 (Sca-1) and myoblast determination protein 1 (MyoD1) were detected by Western-blot. Induction experiments were performed for the pluripotency. Results MDSCs formed multinucleated myotubes in CM and CEECM, and the expression of MyoD1 was up-regulated (P < 0.05). Cells cultured in CKCM were still small round and showed stronger proliferative ability than that in NM (P < 0.05), expressing higher levels of PCNA and Sca-1 (P < 0.05), and expressed muscle marker Desmin, endothelial marker CD34 or glial marker GFAP after multi-directional induction. Conclusions MDSCs mainly undergo myogenic differentiation in NM and CEECM, losing pluripotency. When cultured in CKCM, MDSCs proliferate faster and maintain multi-directional differentiation potential.

Key words: muscle-derived stem cells, different media, proliferation, differentiation