生长板软骨细胞促进骨髓基质干细胞血管内皮生长因子的表达  

Growth plate chondrocytes promoting VEGF expression of BMSCs

作　　者：丁坚[1] 曾炳芳[1] 张先龙[1] 

机构地区：[1]上海交通大学附属第六人民医院骨科,上海市200233

出　　处：《中华创伤骨科杂志》2005年第11期1055-1058,共4页Chinese Journal of Orthopaedic Trauma

基　　金：上海市创伤骨科临床医学中心基金资助课题

摘　　要：目的观察骨髓基质干细胞(BMSCs)在生长板软骨细胞旁分泌作用下血管内皮生长因子(VEGF)的表达规律及其与成骨分化的相关性。方法大鼠BMSCs与生长板软骨细胞进行间接共培养,培养终末期做细胞化学染色,定量测定碱性磷酸酶(ALP)活性,用RT-PCR方法半定量检测VEGFmRNA的表达。结果生长板软骨细胞持续高表达VEGF。BMSCs随共培养时间的延长,ALP活性升高,BMSCs的VEGF的表达也逐渐增强。培养液加入两种分泌型VEGF中和抗体后,VEGF表达趋势不变,ALP活性仍为升高趋势,也不影响培养终末期钙化结节的形成。培养终末期BMSCs的CD31和CD34均阴性。结论BMSCs成骨分化过程中VEGF的表达符合成骨细胞分化基因的表达规律,与成骨细胞特征性基因的表达趋势一致,体外条件共培养条件下,中和VEGF后并不能阻碍BMSCs的成骨分化。Objective To study the regulation of VEGF expression under the paracrine of growth plate chondrocytes and its correlation with osteogenic differentiation of BMSCs. Methods The BMSCs of rats were cocultured with growth plate chondrocytes which were also treated with anti-VEGF as the control group. The BMSCs were evaluated by methods of histochemical stain, ALP （alkaline phosphatase） quantification, and RT-PCR （reverse transcription-polymerase chain reaction） during coeulture. Results The growth plate chondrocytes had high VEGFmRNA expression at different periods during culture. Expression of VEGF mRNA in BMSCs was obviously increased even after addition of anti-VEGF. The tendency was consistent in other gene expressions in osteogenic differentiation. Two endothelial markers （CD31 and CD34） determined by flow cytometry were negative at the end of culture. Conclusion No evidence shows that VEGF is correlated with BMSCs osteogenic differentiation in vitro, but VEGF can be regarded as another marker of osteogenesis.

关 键 词：共培养 血管内皮生长因子 骨髓基质干细胞 生长板软骨细胞 碱性磷酸酶 血管内皮生长因子(VEGF) 骨髓基质干细胞 软骨细胞 生长板 BMSCs 间接共培养 RT-PCR方法 ALP活性 骨细胞分化基因 

分 类 号：R68[医药卫生—骨科学]