【摘要】 [目的]优化骨髓间质干细胞(bone marrow mesenchymal stem cells,BMSCs)体外分离培养技术,改良生物材料聚乙醇酸-乳酸共聚物(poly lactide-co glycolide,PLGA)的细胞黏附性,观察BMSCs与Ⅱ型胶原修饰的PL-GA的黏附性。[方法]密度梯度离心法分离BMSCs,流式细胞分析法(FACS)对细胞表面抗原、细胞活力、细胞周期进行检测,相差显微镜观察细胞形态。相分离法制做PLGA,Ⅱ型胶原进行表面修饰,取第三代干细胞与PLGA附和,扫描电镜观察细胞与材料的黏附情况。[结果]BMSCs可在体外分离扩增,表达CD29、CD44和CD106,不表达CD34和CD45,细胞活力为88.96%,G0-G1细胞占90.32%。细胞形态为长梭形,Ⅱ型胶原表面修饰的PLGA平均孔径为100μm,与BMSCs有较好的黏附性。[结论]BMSCs可在体外长期、稳定培养,是理想的组织工程种子细胞。Ⅱ型胶原表面修饰的PLGA与干细胞有较好的黏附性,可用来做组织工程生物材料。更多还原

【Abstract】 [Objective]To explore a method for isolating and culturing bone marrow mesenchymal stem cell(BMSCs) in vitro,and improve the cellular conglutination ability of biomaterial poly-lactide-co-glycolic acid(PLGA),and observer the adhesion of BMSCs cultured on PLGA modified with Ⅱ collagen.[Method]BMSCs were isolated and cultured in vitro.The cell exterior antigen,cell livingness and cell cycle were analyzed by flow cytometry method,and cellular configuration was observed continually under invertd phase-contrast microscope.PLGA was made by phase separation,and was modified with Ⅱ collagen.The third era BMSCs were cultured on PLGA,and its adhesion with biomaterial was observed scan electron microscope.[Result]BMSCs could be isolated and cultured in vitro,and express CD29,CD44,CD106,but not express CD34,CD45.The cell livingness was 88.96%,cells in G0-Glperiod are 90.32%.The cell morphology was spindle.The average diameter of PLGA modified with Ⅱ collagen was 100 μm,and PLGA has good adhesion with BMSCs.[Conclusion]BMSCs could be cultured stability in vitro for long time,and is good seminal cell of tissue engineering.Ⅱ collagen can improve the cellular adhesion of PLGA,PLGA modified with Ⅱ collagen is the good biomaterial of tissue engineering.更多还原