少突胶质前体细胞分离培养方法及其谱系限定细胞体外分化模型建立的改进

发布时间：2018-07-25 13:35

【摘要】：目的:对少突胶质前体细胞(oligodendrocyte precursor cells,OPCs)的分离、培养及传代方法进行改良,探讨体外条件下OPCs的增殖与分化特性。方法:取新生SD大鼠脊髓来源的细胞分离、纯化OPCs行原代培养,用含碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)和血小板源性生长因子-aa(platelet-derived growth factor-aa,PDGF-aa)的培养液作扩增培养,MTT法检测OPCs的增殖能力;三碘甲腺原氨酸和睫状神经生长因子诱导OPCs分化,最终分化为成熟的少突胶质细胞(oligodendrocytes,OLs),并对分化细胞行形态学及免疫化学染色法鉴定。结果:95%以上的细胞具有双极或三极突起的典型OPCs形态并表达A2B5;MTT法检测显示OPCs在体外保持良好的增殖能力;细胞最终诱导分化为成熟的OLs并表达髓鞘碱性蛋白(myelin basic protein,MBP)。结论:OPCs在体外条件下可继续保持良好的增殖生长及定向分化能力。
[Abstract]:Aim: to improve the method of isolation, culture and passage of oligodendrocyte progenitor cells (oligodendrocyte precursor cells), and to investigate the characteristics of OPCs proliferation and differentiation in vitro. Methods: Spinal cord cells from newborn SD rats were isolated and purified OPCs was cultured in primary culture. The proliferative ability of OPCs was detected by using the culture medium containing basic fibroblast growth factor (basic fibroblast growth factor-b FGF and platelet-derived growth factor-aaaaahuo PDGF-AA, and the differentiation of OPCs was induced by triiodothyronine and ciliary nerve growth factor. Finally, the cells were differentiated into mature oligodendrocytes OLs, and the differentiated cells were identified by morphological and immunocytochemical staining. Results more than 95% of the cells had typical OPCs morphology with bipolar or tripolar protuberances and expressed A2B 5 OPCs. The results showed that OPCs maintained a good proliferative ability in vitro, and the cells eventually differentiated into mature OLs and expressed myelin basic protein (MBP). Conclusion: OPCs can continue to maintain good proliferative growth and directional differentiation ability in vitro.
【作者单位】：中南大学湘雅二医院麻醉科中南大学麻醉医学研究所;
【基金】：国家自然科学基金(81070994,81671962)~~
【分类号】：R741

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