Abstract:

BACKGROUND: Rho kinase inhibition can regulate dynamic reorganization of cytoskeletal proteins, activate transcriptional factor, and stimulate cell proliferation and differentiation.
OBJECTIVE: To explore the effects of Rho kinase inhibitor Y-27632 on proliferation and cell cycle of bone marrow mesenchymal stem cells (BMSCs).
METHODS: Rat BMSCs were isolated using whole bone marrow culture method and purified using adherent culture method in vitro. After two passages, BMSCs were incubated in maintenance medium containing Y-27632 (10 µmol/L, a commonly used working concentration) in 96-well plates. A control group in which BMSCs were not treated was designated.
RESULTS AND CONCLUSION: The growth curves of BMSCs for moderate inoculation density was typically S-shaped. After BMSCs were incubated with Rho kinase inhibitor Y-27632，the lag phase was found during the first 3 days, the growth was accelerated, and the growth curve was upward. Flow cytometry indicated that (80.640±5.516)% of the BMSCs were in G0/G1  phase of cell cycle, (13.397±2.511)% in S phase. After stimulated by Y27632, (61.723±8.829)% of the BMSCs were in G0/G1 phase, and (29.380±7.630) % were in S phase. The cell proliferation during S stage was significantly higher after stimulation by Y27632 (P < 0.05). Rho kinase inhibitor Y-27632 can significantly promote more BMSCs to enter the stage of DNA synthesis, and promote the division and proliferation of BMSCs.