Objective To explore effects of erythropoietin (EPO) on proliferation, activity and cell cycle of rabbit bone marrow mesenchymal stem cells (BMSCs).
Methods A total of four healthy male New Zealand white rabbits aged 6 weeks were supplied by the Experimental Animal Center of Nachang University. Rabbit BMSCs were in vitro isolated by the density gradient centrifugation. Cells were assigned into normal, 2 u/ml, 4 u/ml, 8 u/ml, 16 u/ml groups. In the normal group, cells received normal culture, without additional EPO stimulation. Following EPO intervention, cell morphology and growth were observed. Cell proliferation was detected by MTT assay. Cell cycle changes were measured by flow cytometry. Cell supernatant was collected to detect mass concentration of matrix metalloproteinase-2 (MMP-2).
Results EPO to intervene BMSCs 3d, cells had no abnormal changes, showing colony growth. With increase EPO concentration, cell absorbance gradually increased, especially in the 16 u/ml group. (F=28.029, p=0.008). Compared with the normal group, cell cycle changed in the 2 u/ml, 4 u/ml, 8 u/ml, 16 u/ml groups, and proliferation index significantly increased (p<0.05 or 0.01). Mass concentration of MMP-2 in supernatant was the lowest in the 16 u/ml group, and the highest in the normal group. Significant differences were detected among groups (t=213.21, p<0.001).
Conclusion EPO can increase the proliferative ability of BMSCs.
- cell culture
- bone marrow mesenchymal stem cells
- cell cycle