大鼠颌骨与胫骨骨髓基质细胞生物学特性和基因表达差异的比较  

The Osteogenic Potential of Rat Mandible vs. Tibia Bone Marrow Stromal Cells

作　　者：金辰怡[1] 刘宏伟[1] 

机构地区：[1]同济大学口腔医学院牙周教研室上海牙组织修复与再生工程技术研究中心,上海200072

出　　处：《口腔颌面外科杂志》2016年第3期162-169,共8页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金项目(81271152)

摘　　要：目的 :比较大鼠颌骨来源骨髓基质细胞(M-BMSCs)与胫骨来源骨髓基质细胞(T-BMSCs)生物学特性的差异。方法:体外通过全骨髓贴壁法,分离和培养SD大鼠M-BMSCs和T-BMSCs,取第3代细胞用于:1流式细胞检测鉴定表面标志;2MTT、细胞周期、克隆形成率检测细胞增殖能力差异;3成骨、成脂诱导及染色检测细胞多项分化潜能;4碱性磷酸酶(ALP)活性、染色、免疫荧光染色检测细胞成骨活性的差异;5荧光定量PCR检测Wnt1、Wnt5a、Hoxa11、Runx2、Ocn、Col I的表达差异和成骨诱导后各基因的表达变化。结果:1 M-BMSCs和T-BMSCs阳性表达CD29、CD44,阴性表达CD45;2MTT、细胞周期检测、克隆形成率显示T-BMSCs细胞增殖能力较高;3M-BMSCs在成骨诱导后矿化能力更强;而T-BMSCs成脂诱导形成脂滴的能力更强;4M-BMSCs表达ALP的水平更高;5荧光定量PCR示2种细胞的Wnt1、Wnt5a、Hoxa11、Runx2、Ocn、ColⅠ表达存在差异。结论:体外培养的M-BMSCs和TBMSCs有着不同的生物学特点,其相关基因Wnt1、Wnt5a、Hoxa11、Runx2、Ocn、ColⅠ的表达上存在差异。Objective： To investigate the differences of biological characteristics and expression of related genes between the rat mandible bone marrow stromal cells（M-BMSCs） and tibia bone marrow stromal cells（T-BMSCs） in vitro. Methods：BMSCs were isolated from the SD rat mandible and tibia by bone marrow adherent method in vitro. The passage 3 BMSCs were used to： ①test the markers by flow cytometry, ②test the cell proliferation by MTT and cell cycle and colony form-ing efficiency assay, ③ test the osteogenic and adipogenic differentiation potential by Alizarin red and oil red O stain-ing, ④ test the osteogenic ability by ALP activity staining and immunofluorescence staining, ⑤ test the expression of re-lated genes （Wnt1, Wnt5a, Hoxa11, Runx2, Ocn, Col I） by quantitative real-time PCR after osteogenic induction. Results：①The positive expression of CD29, CD44 and negative expression of CD45 were showed by both M-BMSCs and T-BMSCs.②Stronger ability of proliferation and colony formation was showed by T-BMSCs. ③ Stronger ability of formation of min-eralized nodules was showed by M-BMSCs, stronger ability of formation of lipid droplets was showed by T-BMSCs. ④More ALP were expressed in M-BMSCs. ⑤ Significant differences of expression could be found between related genes （Wnt1, Wnt5a, Hoxa11, Runx2, Ocn, Col I）. Conclusion： Mandible BMSCs have an increased osteogenic potential and augmented capacity to induce bone formation in vitro.

关 键 词：颌骨骨髓基质细胞 胫骨骨髓基质细胞 WNT1 WNT5A HOXA11 

分 类 号：R78[医药卫生—口腔医学]