离心分离法与贴壁分离法对培养骨髓间充质干细胞生物学特性影响.pdfVIP

2010年2月第48卷第5期 ·基础研究 · 离心分离法与贴壁分离法对培养骨髓问充质干细胞生物学特性影响冰 黎 婷 孙晋虎 赵 亮 李 帅 (1．广西医科大学口腔医学院，广西南宁530021；2．广西河池市人民医院，广西河池546300) 摘【要】目的 探讨分离高质量和高活性骨髓间充质干细胞的方法。方法 选取出生2月龄的健康兔，采用密度梯度离心分离 法与贴壁分离法培养骨髓间充质干细胞，检测培养骨髓间充质干细胞生物学特性。结果 贴壁分离法培养的原代骨髓间充 质干细胞贴壁增殖快，细胞活性高，而密度梯度离心分离法培养的原代骨髓间充质干细胞贴壁增殖慢，细胞活性低，但这些 差别在传代后消失。离心分离法得到的细胞纯度较贴壁分离法获得的细胞纯度高。结论 与离心分离法相比贴壁法获得的 原代骨髓间充质干细胞的细胞活性较高、贴壁增殖快，具有方法简单的优点。 [关键词】骨髓；问充质干细胞；兔；细胞培养 【中图分类号】R329．28 文【献标识码]A 文【章编号】1673—9701(2010)05—29—03 Effects of Adherence and Density Gradient Centrifugation Methods on Biological Characteristics of Rabbit Marrow Mesenchymal Stem Cells Cultured inVitro L／Ting1SUNJinhu1ZHAOHan季 uShuaJ1 1．CollegeofStomatology，GuangxiMedicalUniversity，Nanning530021，China；2．Thepeople’sHospitalofHechiCity，Hechi546300，China A【bstract】ObjectiveTostudytheeffectsofadherenceanddensitygradientcentrifugation(DGC)methodsonbiologicalcharacteristicsof rabbitmarrow mesenchymalstem cellsculturedinvitro．Methods Rabbitsaged2monthswereusedtoisolatemarrow mesenchymalstem cells (MSCs)．MSCswithadherenceandDGCmethodswerecultured，passaged，amplifiedandpurifiedinvitro．Thelivingcharacteristicsandadhe— siverateoftheprimaryandgenerationswereobservedrespectively．Thegrowthcurvewasdrawn．Thecellcyclesandcystoskeletonweretestedto studythedifferentMSCsseparatedmehtodsontheproliferationandmetabolism ofMSCs．ResultsMSCswereintactandfibroblastcell-shaped． Adheringspindle-shapedcellswiht highercytoactivewereobservedintheadherenceandDGCseparationrgoups．TheMSCsofprimaryculture inhteadherencergoupshowedmorerapidproliferation，earliercolonyconfluenceandshortertimeforpassagecomparedwithDGCseparationrgoup． Conclusion TheMSCsseparatedbyadherenceshowedarapidproliferation，earliercolonyconfluenceandshortertimeofrthefirstpassage． K【eywords】Marrow；Mesenehymalstemcells；Rabbit；Cellculture 骨髓基质中存在骨髓间充质干细胞 (me