CAJ | 학술논문

目的尝试使用简单的克隆筛选法来纯化骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs)并进行鉴定.方法对早期分离培养的BMSCs,采取低密度接种(10/cm2),待有单克隆形成,用细胞刮除去其他细胞,将克隆的细胞以低密度接种,待再次有单克隆形成时可重复以上操作,反复进行筛选.对筛选的BMSCs生物学特性进行观察,FACs鉴定BMSCs的标记物的表达,并进行成脂、成骨诱导试验.结果采用克隆筛选法培养BMSCs在传代筛选2～3代后即可获得形态均一的细胞群,FACs鉴定细胞高表达BMSCs的阳性标记物(CD29 98.8％,CD90 98.4％),低表达BMSCs的阴性标记物(CD31 2.6％,CD45 3％).BMSCs能被高效地诱导向脂肪和骨分化.结论本实验采用的克隆筛选法可在短期内获得高纯度的BMSCs,方法简单易行而效率高.
목적 상시사용간단적극륭사선법래순화골수간충질간세포(Bone marrow mesenchymal stem cells,BMSCs)병진행감정.방법 대조기분리배양적BMSCs,채취저밀도접충(10/cm2),대유단극륭형성,용세포괄제거기타세포,장극륭적세포이저밀도접충,대재차유단극륭형성시가중복이상조작,반복진행사선.대사선적BMSCs생물학특성진행관찰,FACs감정BMSCs적표기물적표체,병진행성지、성골유도시험.결과 채용극륭사선법배양BMSCs재전대사선2～3대후즉가획득형태균일적세포군,FACs감정세포고표체BMSCs적양성표기물(CD29 98.8％,CD90 98.4％),저표체BMSCs적음성표기물(CD31 2.6％,CD45 3％).BMSCs능피고효지유도향지방화골분화.결론본실험채용적극륭사선법가재단기내획득고순도적BMSCs,방법간단역행이효솔고.
Objective To purify BMSCs by a simple Colony-Forming system and identify BMSCs in vitro.Methods BMSCs were planted at low-density ( 10/cm2 ) in the early stage of isolation,additional cells were scraped until a colony formed,secondly seeded those Colony-Forming cells at low-density.This process was repeat again when passaged cells formed another colony.This so called Colony-Forming selection was repeated several times until highly purified cells were obtained.Biological characters of BMSCs were observed.The surface antigens of BMSCs were identified by flow cytometry.The multipotentiality of BMSCs was assayed for differentiation into either osteoblasts or adipocytes.Results Homogeneous cells were obtained after BMSCs were passaged twice to thrice by Colony-Forming system.These BMSCs highly expressed positive surface antigens of BMSCs ( CD29 in 98.8％,CD90 in 98.4％ ) meanwhile seldom expressed negative surface antigens of BMSCs (CD31 in 2.6％,CD45 in 3％ ).BMSCs efficiently differentiated into osteoblasts and adipocytes.Conclusions Colony-Forming system is a simple and effective way to get highly purified BMSCs in a short time.