犬颌骨与髂骨两种骨髓间充质干细胞增殖和骨向分化能力比较  

Comparison of the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells from canine jaw and iliac

作　　者：王国轩[1] 朱国雄[2] 朱斌[3] 梁世桢[1] 赵喜聪[3] 安玉林[3] 金岩[3] 

机构地区：[1]辽宁医学院济南军区总医院研究生培养基地,山东济南250031 [2]济南军区总医院口腔科,山东济南250031 [3]第四军医大学口腔医学院,陕西西安710032

出　　处：《牙体牙髓牙周病学杂志》2012年第11期623-627,632,共6页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金重点项目(31030033)

摘　　要：目的:比较犬颌骨与髂骨两种骨髓间充质干细胞增殖和骨向分化能力,为骨移植和组织工程骨的种子细胞获取提供理论基础。方法:体外组织块培养法和有限稀释法克隆化培养颌骨骨髓间充质干细胞(J-BMMSCs组);骨髓穿刺法和密度梯度离心法培养髂骨骨髓间充质干细胞(I-BMMSCs组)。取第3代细胞分别进行以下检测:①流式细胞仪鉴定细胞表面抗原标记;②计算克隆形成率;③MTT法检测生长曲线;④骨向诱导和RT-PCR检测成骨相关基因Runx-2、BSP mRNA表达情况。结果:①J-BMMSCs组和I-BMMSCs组均阳性表达CD44、CD29,阴性表达CD34;②J-BMMSCs组和I-BMMSCs组的克隆形成率分别为(24.5±2.3)%和(18.6±1.3)%(P<0.05);③MTT法检测生长曲线显示J-BMMSCs组增殖速度强于I-BMMSCs组(P<0.05);④成骨诱导7 d后,ALP染色结果显示J-BMMSCs组的成骨能力强于I-BMMSCs组;成骨诱导21 d后两组均出现不同程度矿化结节,定量分析显示J-BMMSCs组矿化能力强于I-BMMSCs组;成骨诱导0 d、7 d、14 d后,RT-PCR结果显示J-BMMSCs组各时间点Runx-2和BSP mRNA表达均明显高于I-BMMSCs组。结论:J-BMMSCs组增殖及成骨能力均强于I-BMMSCs组。颌骨是骨移植的理想供体位点,颌骨骨髓适于用作组织工程骨的成体干细胞来源。AIM ： To investigate the proliferation and osteogenic differentiation potential of canine bone mar- row mesenchymal stem cells from jaw（J-BMMSCs） and iliac （I-BMMSCs））, so as to provide an experimental evidence for choosing a good donor site for bone graft and a new source of seed cells in bone tissue engineering. METHODS： J-BMMSCs were isolated and cultured by limited dilution of culture cells for single cell clone. I-BMMSCs were taken by puncture and isolated by density gradient centrifugation. Passage 3 cells were collected and the surface markers of the cells were detected with flow cytometry. Cloning efficiency and growth curves were used to investigate the prolifera- tion of J-BMMSCs and I - BMMSCs. The osteogenic differentiation capacity of the two BMMSC groups was evaluated by ALP, alizarin red staining and real time PCR. RESULTS ： The expression of cell surface molecules CD44 and CD29 of the two BMMSCs was all positive and that of CD34 was all negative. Clone formation rates of J-BMMSCs and I - BMMSCs were 24.5% and 18.6%, respectively （P 〈 0.05 ）. The proliferation rate of J-BMMSCs was significantly higher than that of I-BMMSCs by MTY assay. Alizarin red staining showed varying degree of mineralized nodules after 21-day induction,the expression of osteogenic genes Runx-2 and BSP of J-BMMSCs was evidently higher than that of I-BMMSCs after 0 d,7 d and 14 d induction（P 〈 0.05 ）. CONCLUSION： The proliferation and osteogenic differentiation capacity of J-BMMSCs are significantly higher than those of I-BMMSCs. Jaw is a prime donor site for bone graft and an important source of seed cells in bone tissue engineering.

关 键 词：骨髓间充质干细胞 颌骨 髂骨 骨向分化 

分 类 号：R780.2[医药卫生—口腔医学]