贵州小型猪骨髓间充质干细胞的分化潜能分析

Qian Li, Zengcai Liang, Taofeng Lu
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摘要

本研究利用造血干细胞和骨髓间充质干细胞的不同生长特性,采用全骨髓粘附法分离培养BMSCs,并用RT-PCR检测细胞表面标志物CD29、CD31、CD34、CD44、CD71、CD73、CD90。体外诱导成骨细胞分化为脂肪细胞和成骨细胞,并用油红 O 和鲁宾宁染色观察其分化能力。研究贵州小型猪(Sussa lvanius)BMSCs的体外分离和培养方法,并研究其生物学特性和多元分化潜力。结果表明,体外培养的原代BMSCs在48小时内可见少量粘附细胞,5~6天后汇合细胞达到90%。细胞生长状态良好,呈纺锤形和多角形。RT-PCR 检测细胞表面 CD29、CD44、CD71、CD73 和 CD90 阳性表达,CD31 和 CD34 阴性表达。油红 O 染色显示细胞内有脂滴形成,茜素红染色显示细胞内有钙结节,表明本实验分离培养的细胞具有多向分化能力。这符合 BMSCs 的特征。体外培养的 BMSCs 生长稳定,扩增后的 BMSCs 具有多向分化潜能,是理想的组织工程种子细胞。
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Differentiation Potential Analysis of Bone Marrow Mesenchymal Stem Cells from Guizhou Miniature Pigs
In this study, different growth characteristics of hematopoietic stem cells and bone marrow mesenchymal stem cells were used to isolate and culture BMSCs by whole bone marrow adhesion method, and RT-PCR was used to detect cell surface markers CD29, CD31, CD34, CD44, CD71, CD73, CD90. The differentiation of osteocytes into adipocytes and osteoblasts was induced in vitro, and the differentiation ability was observed by staining with oil red O and rubinin. To investigate the isolation and culture methods of BMSCs from Guizhou miniature pigs (Sussa lvanius) in vitro, and to study their biological characteristics and potential for multi-differentiation. The results showed that a small number of adherent cells could be seen in the primary BMSCs cultured in vitro within 48 hours, and the confluent cells reached 90% in 5~6 days. The cell growth state was good, and the shape was spindle and polygonal. RT-PCR was used to detect the positive expression of CD29, CD44, CD71, CD73 and CD90 on the cell surface, and the negative expression of CD31 and CD34. Oil red O staining showed the formation of lipid droplets in the cells, and alizarine red staining showed calcium nodules in the cells, indicating that the cells isolated and cultured in this experiment had the ability of multidirection differentiation. It is consistent with the characteristics of BMSCs. The growth of BMSCs cultured in vitro was stable, and the expanded BMSCs had multidirectional differentiation potential, which was an ideal seed cell for tissue engineering.
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