CAJ | 학술논문

【目的】建立兔骨髓间充质干细胞（MSCs）的分离、纯化及鉴定方法，观察体外成骨潜能。【方法】应用密度梯度离心联合贴壁筛选的方法，体外分离培养兔骨髓来源的M SCs ，培养过程中倒置显微镜下观察其生物学特性，流式细胞仪检测细胞表型，所得的细胞进行成骨诱导分化后进行碱性磷酸酶（alkaline phos‐phatase ，ALP）染色及茜素红染色鉴定，并对成骨分化指标进行检测。【结果】原代分离培养的贴壁细胞呈长梭形，漩涡状排列。传代后增殖迅速，细胞为单一的梭形，排列更加有序。培养的细胞CD44呈阳性表达，而CD34呈阴性表达。成骨诱导后ALP染色和茜素红染色均呈阳性。诱导组成骨分化后成骨标志物含量较对照组明显要高。【结论】密度梯度离心结合贴壁的方法可以获得高纯度MSCs ，增殖旺盛，体外具有成骨潜能。
【목적】건립토골수간충질간세포（MSCs）적분리、순화급감정방법，관찰체외성골잠능。【방법】응용밀도제도리심연합첩벽사선적방법，체외분리배양토골수래원적M SCs ，배양과정중도치현미경하관찰기생물학특성，류식세포의검측세포표형，소득적세포진행성골유도분화후진행감성린산매（alkaline phos‐phatase ，ALP）염색급천소홍염색감정，병대성골분화지표진행검측。【결과】원대분리배양적첩벽세포정장사형，선와상배렬。전대후증식신속，세포위단일적사형，배렬경가유서。배양적세포CD44정양성표체，이CD34정음성표체。성골유도후ALP염색화천소홍염색균정양성。유도조성골분화후성골표지물함량교대조조명현요고。【결론】밀도제도리심결합첩벽적방법가이획득고순도MSCs ，증식왕성，체외구유성골잠능。
[Objective] To establish the experimental method for isolating and culturing rabbit bone‐de‐rived mesenchymal stem cells (MSCs) and examine their osteogenic potentials in vitro .[Methods]Mesenchy‐mal stem cells (MSCs) were isolated and purified by density gradient centrifugation plus attachment culture . The biological characteristics of MSCs were observed under inverted microscope .Cell surface markers were as‐sessed by flow cytometry and MSCs identified with alkaline phosphatase (ALP) and Alizarin red staining after osteogenic differentiation .Finally the markers of osteogenic differentiation were detected .[Results] The ad‐hered cells by primary culture were spindle and whirlpool‐shaped .After passage ,the cells became spindle‐shaped and distributed orderly .Cultured cells were positive for CD44 ,but negative for CD34 .After osteogenic induction ,ALP and Alizarin red stains were positive .After induced osteogenic differentiation ,the content of osteogenic markers in osteogenic differentiation group was significantly higher than that of control group .[Conclusion] Highly purified MSCs may be obtained by density gradient separation and screening adherence . And MSCs possess osteogenic potential in vitro .