骨髓基质干细胞诱导内皮细胞与自体骨髓基质干细胞低氧条件下培养后的成骨能力  

Osteogenic character of autologous bone marrow stromal cells cocultured under hypoxia with endothelial cells induced by bone marrow stromal cells

作　　者：曲伟栋 杜平功 赵华强[2] 

机构地区：[1]烟台市口腔医院口腔颌面外科,山东烟台264000 [2]山东大学口腔医院.口腔医学院口腔颌面外科,山东济南250012

出　　处：《中国口腔颌面外科杂志》2011年第6期455-460,共6页China Journal of Oral and Maxillofacial Surgery

摘　　要：目的:探讨骨髓基质干细胞(BMSCs)诱导内皮细胞(ECs)与自体BMSCs低氧条件下联合培养对BMSCs成骨能力的影响。方法:体外分离培养兔BMSCs并向ECs诱导培养,用CD34免疫细胞化学染色鉴定内皮细胞表型;实验分组为BMSCs常氧成骨诱导培养组,BMSCs低氧成骨诱导培养组,BMSCs+ECs常氧成骨诱导联合培养组,BMSCs+ECs低氧成骨诱导联合培养组,持续培养7d,前6d应用PNPP法每天固定时间测定碱性磷酸酶(ALP)活性并作统计学分析;茜素红染色观察第7d矿化结节形成情况。低氧组为1.0%O2浓度。采用SPSS11.0软件包对数据进行方差分析和q检验。结果:BMSCs在一定诱导条件下可诱导为ECs,CD34免疫细胞化学染色为阳性;统计学分析表明,BMSCs+ECs低氧成骨诱导联合培养组的ALP活性表达显著高于其他3组(P<0.05)。只有BMSCs+ECs低氧成骨诱导联合培养组有钙化结节形成,茜素红染色呈橘红色。结论:在一定时间内,由BMSCs诱导来源的ECs与自体BM-SCs低氧条件下联合培养,BMSCs的成骨活性可显著提高。PURPOSE： To investigate the influence of hypoxia and co-culture of endothelial cells（ECs） induced by BMSCs and autologous bone marrow stromal cells（BMSCs） on proliferation and osteogenetic ability of BMSCs.METHODS：The rabbit BMSCs were isolated and induced into endothelial cells.Then induced endothelial cells were identified by CD34 immunocytochemistry.The BMSCs and ECs were divided into four groups.（1） BMSCs group cultured in osteogenic medium under normoxia;（2） BMSCs group cultured in osteogenic medium under hypoxia;（3） BMSCs＋ECs group cocultured in osteogenic medium under normoxia;（4） BMSCs＋EC group cocultured in osteogenic medium under hypoxia.Four groups were cultured for 7 days in vitro and alkaline phosphatase（ALP） activity was detected by PNPP assay once a day within 6 days.Then the ALP activity curves were drawn according to OD values.Mineralization nodules of four groups were dyed by alizarin monosulfonate after 7 days＇ cultivation.The data was analyzed using SPSS11.0 software package.RESULTS： The rabbit BMSCs can be induced into ECs in vitro.The ALPactivity of BMSCs＋ECs group cocultured in osteogenic medium under hypoxia was the strongest in four groups（P0.05）.The mineralized nodules were formed only in this group and dying was positive.There was no mineralized nodule formed in other groups.CONCLUSION： Autologous BMSCs cocultured under hypoxia with endothelial cells induced by BMSCs can significantly increase the proliferating ability of BMSCs.

关 键 词：骨髓基质干细胞 内皮细胞 低氧 联合培养 体外成骨 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]