CAJ | 학술논문

目的探讨有效的分离与纯化大鼠骨髓间充质干细胞（BMSCs）方案，并研究BMSCs基本生物学特性。方法取SPF级4周龄健康SD大鼠1只，采用改良全骨髓贴壁法分离提取BMSCs，用倒置相差显微镜观察细胞形态变化及其生长特性，利用流式细胞仪检测P3代细胞表面标志物，细胞计数试剂盒（CCK）-8法绘制P3代细胞生长曲线。结果体外分离的原代细胞可见典型的贴壁生长，形态以梭形和多角形为主，呈旋涡状细胞集落，约8 d可达80%以上融合。传代纯化后细胞形态均一，具有较强的增殖能力。生长曲线提示BMSCs呈S形趋势。P3代 BMSCs代表面标志物分化群（CD）29、CD90阳性表达，而CD11b、CD45阴性表达。结论改良全骨髓贴壁法可获得高纯度、增殖活跃、生物学特性稳定的BMSCs，BMSCs的生长曲线呈S形趋势。
목적탐토유효적분리여순화대서골수간충질간세포（BMSCs）방안，병연구BMSCs기본생물학특성。방법취SPF급4주령건강SD대서1지，채용개량전골수첩벽법분리제취BMSCs，용도치상차현미경관찰세포형태변화급기생장특성，이용류식세포의검측P3대세포표면표지물，세포계수시제합（CCK）-8법회제P3대세포생장곡선。결과체외분리적원대세포가견전형적첩벽생장，형태이사형화다각형위주，정선와상세포집락，약8 d가체80%이상융합。전대순화후세포형태균일，구유교강적증식능력。생장곡선제시BMSCs정S형추세。P3대 BMSCs대표면표지물분화군（CD）29、CD90양성표체，이CD11b、CD45음성표체。결론개량전골수첩벽법가획득고순도、증식활약、생물학특성은정적BMSCs，BMSCs적생장곡선정S형추세。
ObjectiveTo explore an effective method for the isolation and purification of rats bone marrow mesenchymal stem cells (BMSCs) and to study the basic biological characteristics of BMSCs. MethodsOne healthy Sprague-Dawley (SD) rat of specific pathogen free (SPF) grade aged 4 weeks was selected, and the BMSCs were isolated using the modiifed whole bone marrow adherent method. The morphologic variation and proliferation features of BMSCs were observed under inverted phase contrast microscope, and the surface markers of BMSCs of P3 generation were detected by flow cytometry. Cell growth curves were drawn using the cell counting kit (CCK)-8 assay.ResultsTypical adherent growth was observed in the primary BMSCs with the forms of fusiform or polygon and appearance of circinate cell colony. The cell colonies fused with each other up to 80% in about 8 d. The puriifed passage cells still kept a homogeneous morphology and a high potential of proliferation. The growth curve of BMSCs indicated an S shape. The surface markers of BMSCs of P3 generation cluster of differentiation (CD) 29 and CD90 were observed positive, but CD11b and CD45 negative. ConclusionsBMSCs with high-purity, active proliferation and stable biological characteristics can be cultured using modiifed whole bone marrow adherent method. The growth curve of BMSCs presents in an S shape.