大鼠颌骨与胫骨骨髓基质细胞间接共培养后生物学差异的比较  

Comparison of biological difference between rat mandible and tibia bone marrow stromal cells after Indirect co-culture

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,200072

出　　处：《现代口腔医学杂志》2016年第4期198-203,共6页Journal of Modern Stomatology

基　　金：国家自然科学基金项目(81271152)

摘　　要：目的 SD大鼠颌骨骨髓基质细胞(M-BMSCs)与胫骨骨髓基质细胞(T-BMSCs)体外间接共培养后,检测两者生物学特性和相关基因表达的变化。方法应用全骨髓贴壁法,分离和培养M-BMSCs和T-BMSCs。取第三代细胞用Transwell小室间接共培养后,做以下检测。1MTT检测细胞增殖能力差异;2ALP活性、ALP染色、免疫荧光染色检测细胞成骨活性的差异;3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的表达变化。结果 1全骨髓贴壁法能分离和培养M-BMSCs和T-BMSCs;2间接共培养后,M-BMSCs和T-BMSCs增殖活力减弱;3共培养条件提高了T-BMSCs和抑制M-BMSCs早期表达ALP的能力;4间接共培养后,M-BMSCs和TBMSCs中Wnt1、Wnt5a、Hoxa11、Runx2、Ocn、Col I基因表达出现变化。结论 M-BMSCs和T-BMSCs间接共培养后,两者的生物学特点的改变和相关基因表达改变。Objective To investigate the biological difference and variation of related genes expression between SD rat mandible bone marrow stromal cells（M-BMSCs）and tibia bone marrow stromal cells（T-BMSCs）after indirect co-culture in vitro. Methods The BMSCs were isolated from the mandible and tibia by whole bone marrow adherent culture in vitro. These two BMSCs of passage 3 were co-cultured by Transwell system and then tested as follows. 1The cell proliferation was tested by MTT assay; 2 The osteogenic ability was tested by ALP activity, ALP staining and immunofluorescence staining; 3The expression of related genes, which contained Wnt1, Wnt5 a, Hoxa11, Runx2, Ocn and Col I, were tested by Quantitative Real-time PCR. Results 1Both M-BMSCs and T-BMSCs could be cultured by whole bone marrow adherent method in vitro; 2 The proliferation ability of both cells weakened after indirect co-culture after indirect co-culture; 3ALP activity was Increased in T-BMSCs while decreased in M-BMSCs after indirect co-culture; 4The expression of related genes（Wnt1, Wnt5 a, Hoxa11, Runx2, Ocn, Col I）in both cells varied after indirect co-culture.Conclusion Analysis of these results demonstrated significant biological difference and variation of related genes expression between M-BMSCs and T-BMSCs after indirect co-culture in vitro.

关 键 词：颌骨骨髓基质细胞 胫骨骨髓基质细胞 间接共培养 WNT1 WNT5A HOXA11 

分 类 号：R782[医药卫生—口腔医学]