骨髓间质干细胞与Ⅱ型胶原修饰的 PLGA黏附性的观察_医学论文.docVIP

骨髓间质干细胞与Ⅱ型胶原修饰的 PLGA黏附性的观察_医学论文 骨髓间质干细胞与Ⅱ型胶原修饰的 PLGA黏附性的观察_医学论文 【摘要】 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，PLGA)的细胞黏附性，观察BMSCs与Ⅱ型胶原修饰的PLGA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100 μm，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。 【关键词】 骨髓间质干细胞 PLGA 组织工程 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 G0hesion 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. Key words：bone marrow mesenchymal stem cell PLGA tissue engineering 　　 组织工程修复受损组织的基本方法就是将种子细胞种植于生物支架材料上，经体外培养一定时间后再植入体内达到修复和重建受损组织的目的，所以细胞与生物支架材料的黏附性就成了构建组织工程化组织和器官的关键问题之一。本研究将体外提纯，扩增的BMSCs种植到经Ⅱ型胶原修饰的PLGA材料上联合培养，扫描电镜观察其黏附情况，观察Ⅱ型胶原修饰的PLGA材料的细胞亲和性，探索以BMSC为种子细胞，以Ⅱ型胶原修饰的PLGA为生物支架材料构建组织工程的可行性，为临床组织工程的应用研究提供理论和实验数据。 1 材料和方法