共培养大鼠内皮祖细胞对自体骨髓基质干细胞成骨作用的影响  

Influence of endothelial progenitor cells on osteogenic effects of autologous bone marrow stromal cells of rats in vitro co-culture condition

作　　者：韦惠平[1] 庞芳河[1] 

机构地区：[1]广西医科大学口腔医学院,南宁530021

出　　处：《中国临床新医学》2011年第5期424-429,共6页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

摘　　要：目的探讨体外培养大鼠外周血内皮前体细胞(EPCs)与自体骨髓基质细胞(BMSCs)共培养时对BMSCs成骨作用的影响。方法采用密度梯度离心法分离、培养大鼠BMSCs和EPCs,培养细胞分为四组:A组(BMSCs组)、B组(EPCs组)、C组(BMSCs成骨诱导组)及D组(BMSCs和EPCs联合培养组)。通过观察细胞克隆形态、免疫细胞化学、细胞增殖、碱性磷酸酶活性,从酶学、组织学及生化等不同方面观察:EPCs对BMSCs成骨活性及生长情况的影响。结果免疫细胞化学染色证实C组培养的细胞具有晚期EPCs的特性。倒置相差显微镜、HE染色均证实共培养的BMSCs和EPCs生长良好,并能够形成与单纯成骨诱导培养的BMSCs相似的钙结节。MTT检测结果:各组细胞增殖差异无统计学意义(P>0.05)。碱性磷酸酶活性检测结果:C、D组显著高于A、B组(P<0.05)。结论 EPCs和BMSCs联合培养具有良好的细胞相容性,EPCs能够增强成骨细胞的ALP活性,提高成骨细胞的增殖能力。Objective To observe the influence of endothelial progenitor cells （EPCs） on the osteogenic effects of autologous bone marrow stromal cells （BMSCs） of rats in vitro co-culture of EPCs and BMSCs. Methods BMSCs and EPCs were obtained from rat bone marrow and peripheral blood by density gradient centrifugation. The cultured ceils were divided into four groups：group A （BMSCs）, group B （EPCs osteogenic induction）, group C （BM- SCs osteogenic induction） and group D（co-culture of induced BMSCs and EPCs）. The EPCs were identified with immunocytochemistry staining. The ceil proliferation and alkaline phosphatase （ALP） activity were observed to determine the effects of the osteogenic potential and cellular compatibility in each group. Results The cells were able to line up in the cord-like structure and showed CD31 ,vWF,positive in group B. There was good compatibility when EPCs and induced BMSCs were co-cultured. The activity of ALP in the group C and D was higher than the other two groups. Conclusion In direct co-culture system in vitro, the cellular compatibility is perfect. EPCs can improve ALP activity of induced osteoblasts.

关 键 词：组织工程 骨髓基质细胞 内皮前体细胞 联合培养 

分 类 号：R78[医药卫生—口腔医学]