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

doi:10.3969/j.issn.1005-4979.2016.03.003

《口腔颌面外科杂志》 ›› 2016, Vol. 26 ›› Issue (3): 162-169. doi: 10.3969/j.issn.1005-4979.2016.03.003

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

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

收稿日期:2016-01-21 修回日期:2016-02-24 出版日期:2016-06-28 发布日期:2016-08-08
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）

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

JIN Chen-yi, LIU Hong-wei

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

摘要/Abstract

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

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

中图分类号:

R782
Q813.1

金辰怡，刘宏伟. 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[J]. 《口腔颌面外科杂志》, 2016, 26(3): 162-169.

JIN Chen-yi, LIU Hong-wei. The Osteogenic Potential of Rat Mandible vs. Tibia Bone Marrow Stromal Cells[J]. 《Journal of Oral and Maxillofacial Surgery》, 2016, 26(3): 162-169.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2016/V26/I3/162

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