《Journal of Oral and Maxillofacial Surgery》 ›› 2016, Vol. 26 ›› Issue (3): 162-169. doi: 10.3969/j.issn.1005-4979.2016.03.003

• Basic Scientific Study • Previous Articles     Next Articles

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

JIN Chen-yi, LIU Hong-wei   

  1. Department of Periodontics, School of Stomatology, Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China
  • Received:2016-01-21 Revised:2016-02-24 Online:2016-06-28 Published:2016-08-08

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

金辰怡,刘宏伟   

  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
  • 通讯作者: 刘宏伟,教授. E-mail:hwliu@tongji.edu.cn
  • 作者简介:金辰怡(1988—),女,浙江嘉兴人,住院医师,硕士. E-mail: judyhzdn@163.com
  • 基金资助:

    国家自然科学基金项目(81271152)

Abstract: 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: ① test the markers by flow cytometry, ② test the cell proliferation by MTT and cell cycle and colony forming efficiency assay, ③ test the osteogenic and adipogenic differentiation potential by Alizarin red and oil red O staining, ④ test the osteogenic ability by ALP activity staining and immunofluorescence staining, ⑤ test the expression of related genes (Wnt1, Wnt5a, Hoxa11, Runx2, Ocn, Col I) by quantitative real-time PCR after osteogenic induction. Results: ① The positive expression of CD29, CD44 and negative expression of CD45 were showed by both M-BMSCs and T-BMSCs. ② Stronger ability of proliferation and colony formation was showed by T-BMSCs. ③ Stronger ability of formation of mineralized nodules was showed by M-BMSCs, stronger ability of formation of lipid droplets was showed by T-BMSCs. ④ More 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.

Key words: mandible bone marrow stromal cells(M-BMSCs), tibia bone marrow stromal cells(T-BMSCs), Wnt1, Wnt5a, Hoxa11

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、Col I表达存在差异。结论:体外培养的M-BMSCs和T-BMSCs有着不同的生物学特点,其相关基因Wnt1、Wnt5a、Hoxa11、Runx2、Ocn、Col I的表达上存在差异。

关键词: 颌骨骨髓基质细胞;  , 胫骨骨髓基质细胞;  , Wnt1;  , Wnt5a;  , Hoxa11

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