骨髓间充质干细胞与成骨细胞复合培养对成骨细胞增殖的影响

Effect of co-culture of bone-marrow mesenchymal stem cells and osteoblasts on proliferation of osteoblasts

目的探讨骨髓间充质干细胞(BMSCs)与成骨细胞复合培养对成骨细胞增殖的影响。方法将BMSCs与成骨细胞分别按1∶2、1∶4、1∶1、2∶1和4∶1比例复合培养(分别为A1组、A2组、A3组、A4组和A5组);另设成骨细胞培养组(B1组)和BMSCs培养组(B2组)为对照。流式细胞术检测复合培养组的细胞比例;细胞计数试剂盒8(CCK-8)测定所有细胞培养组的生长曲线;RT-PCR检测碱性磷酸酶(ALP)及Ⅰ型胶原mRNA表达。结果 BMSCs与成骨细胞复合培养能促进成骨细胞增殖,以A2组效果最好,明显大于B1组(P<0.05);而A2组ALP及Ⅰ型胶原mRNA表达与B1组相比差异无统计学意义(P>0.05)。结论 BMSCs与成骨细胞复合培养能促进成骨细胞增殖,其中1∶4比例最佳。

Objective To investigate the effect of co-culture of bone-marrow mesenchymal stem cells（BMSCs） and osteoblasts on proliferation of osteoblasts.Methods BMSCs and osteoblasts were co-cultured according to different ratios of 1∶2（group A1）,1∶4（group A2）,1∶1（group A3）,2∶1（group A4） and 4∶1（group A5）.Culture of osteoblasts alone（group B1） or BMSCs（group B2） were taken as the controls.The ratios of the co-cultured cells were detected by flow cytometry.The growth curve of each group was determined by cell counting kit-8（CCK-8）,and the mRNA expressions of alkaline phosphatase（ALP） and collagen type I were measured by RT-PCR.Results Co-culture of BMSCs and osteoblasts promoted the proliferation of osteoblasts,especially in group A2,which was obviously greater than that in group B1（P0.05）.But there was no significant difference in mRNA expressions of ALP and collagen type I between groups of A2 and B1（P0.05）.Conclusion Co-culture of BMSCs and osteoblasts can enhance the proliferation of osteoblasts,for which the best ratio of co-culture is 1∶4.