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目的:探讨兔骨髓间充质干细胞(r BMMSCs)膜片的基本生物学特性及其在组织工程应用中的优势。方法:从日本大耳白兔提取原代骨髓培养r BMMSCs,采用简单制备方法构建r BMMSCs细胞膜片,大体、HE染色观察,MTT、成骨成脂诱导检测膜片增殖分化能力,RT-PCR检测膜片相关优势基因的表达情况。结果:成功分离r BMMSCs,诱导10-14d后获得白色膜状细胞膜片,并可用无菌眼科镊提拉。镜下观察细胞呈梭形重叠生长,HE染色显示细胞间紧密连接并有大量细胞外基质(ECM)分泌;MTT显示膜片细胞密度随时间增加;成骨成脂诱导结果显示其分化能力明显优于普通培养的r BMMSCs;RT-PCR结果显示,与r BMMSCs相比,细胞膜片高表达ColⅠ和Fn。结论:r BMMSCs细胞膜片相比普通培养的r BMMSCs,在组织工程应用中更具优势。
Objective: To investigate the basic biological characteristics of rabbit bone marrow mesenchymal stem cells (r BMMSCs) and their advantages in tissue engineering application. Methods: Primary BMMSCs were isolated from Japanese white rabbits and r BMMSCs were cultured by simple preparation method. The morphological changes of the cells were observed by HE staining, MTT and osteogenic adipogenic induction. RT-PCR Diaphragm related advantages of gene expression. Results: The r BMMSCs were successfully isolated and the white membranous membranous membrane was obtained after 10-14 days of induction and could be pulled by sterile ophthalmic forceps. Microscopically, the cells showed spindle-shaped overlapping growth. HE staining showed that the cells were tightly connected and had a large amount of extracellular matrix (ECM) secretion. MTT showed that the cell density of the membrane increased with time; osteogenic and adipogenic induction showed that the differentiation ability was obvious Compared with r BMMSCs, the expression of Col I and Fn was highly expressed in the membrane of r BMMSCs. Conclusion: BMMSCs r BMMSCs compared with the normal culture r BMMSCs, tissue engineering more advantages.