The Effect Of Cytokines Pretreatment On The Proliferation And The Differentiation Of Bone Mesenchymal Stem Cells Into Cardiomyogenic Cells

Objective: To investigate the effect of Stem Cells Factor(SCF) and Granulocyte Colony-Stimulating Factor(G-CSF) pretreatment on the proliferation and the differentiation of MSCs into cardiomyogenic cells.Methods: Forty eight SD rats were divided randomly into four groups. The control group: sham treatment; the group SCF: the rats were treated with SCF; the group G-CSF: the rats were treated with G-CSF; the group SCF+G-CSF: the rats were treated with SCF and G-CSF together. After treated with SCF(20 μg/kg) and G-CSF(10μg/kg) every day about 5 days, the MSCs were isolated primarily from bone marrow of rat and purified by passage culture. The 4th passage of MSCs were labled with DAPI and detected by Flow Cytometry before co-culture. Meanwhile, the activity of cardiomyoc-ytes were also detected before co-culture. The neonatal rat cardiomyocytes were co-cultured with DAPI-MSCs after 3 days. And Troponin T was also examined by immunocytochemical method in the 3th day. We analyse the percentage of the differentiation of MSCs into cardiomyogenic cells during the co-culture days. The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were detected respectively with digital microscope camera system and immunofluorescence technique.Results: Compared with group SCF, G-CSF and the control group, the MSCs were markedly induced into S phase after treated with SCF and G-CSF together. Meanwhile, the percentage of MSCs in S phase from group SCF, G-CSF were significantly higher than from the control group. Moreover, The percentage of MHC proteins-positive MSCs and TnT proteins-positive MSCs in SCF+G-CSF group were significantly higher than that in group SCF, G-CSF and the control group. And the percentage of MHC proteins-positive MSCs and TnT proteins-positive MSCs in group SCF and group G-CSF were also markedly higher than that in control group.Conclusion: SCF and G-CSF stimulated the proliferation of MSCs and induced MSCs to differentiate into cardiomyocytes distinctly, and it was more effective by using SCF and G-CSF together.