| Objective: To investigate the effect of VEGF and PDGF-BB on proliferation of bone marrow mesenchymal stem cells(BMSCs)and mRNA expression levels of related gene indicators at different stages of angiogenesis,to provide theoretical basis for optimizing tissue-engineered stem cells.Methods: Primary bone marrow mesenchymal stem cells(BMSCs)were obtained from SD rat and cultured in vitro.The cells were treated with VEGF,PDGF-BB and VEGF + PDGF-BB respectively.With serial concentration of VEGF(0,20,40,60,80,100,120 ng/mL)and PDGF-BB(0,20,40,60,80,100,120 ng/mL)for seven consecutive days to test cell proliferation capability and determine the optimal concentration for promoting cell proliferation.RNA were extracted after 7 and 14 days of treatment,and the mRNA expression levels of Ang-1,Hif-1α,HGF,IGF and other related vascular genes were detected by RT-PCR.Results:(1)Proliferation: after the addition of growth factors,cell proliferation was significantly improved,and the effect of VEGF or PDGF-BB alone was not as good as that of combined treatment.The optimal combination concentration was 80 ng/mL VEGF + 80 ng/mL PDGF-BB.(2)mRNA expression levels of Ang-1,Hif-1α,HGF and IGF gene indicators were significantly up-regulated compared the control group(p < 0.05).Among them,VEGF+PDGF-BB had the most significant effect on promoting BMSCs angiogenesis(p < 0.05).(3)Changes in mRNA expression of vascularized genes.The mRNA expression levels of Hif-1α,HGF,Ang-1 and IGF in the control group were all lower at day 14 than at day 7.Compared to control group,the expression of Hif-1α and HGF in the group treated with VEGF or PDGF-BB alone or in combination was higher at day 14 than at day 7,while the expression of Ang-1 and IGF was lower at day 14 than at day 7..Conclusion: In vitro experiments showed that the combined application of VEGF and PDGF-BB could not only promote the proliferation of SD rat BMSCs but also promote the expression of vascular genes. |
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