彩图骨髓干细胞体外组织工程软骨形成实验探究.docVIP

彩图骨髓干细胞体外组织工程软骨形成实验探究 　 作者：宋世锋，肖海涛，武伟，袁素，谢瑶云，吴多能，白殿卿 【摘要】 [目的]探讨骨髓干细胞(BMSCs)在体外不同载体中软骨形成的能力和可行性。[方法]分离培养猪BMSCs，将第3代BMSCs以载体内多点注射和表面点滴法植于A组：Ⅱ型纤维胶原蛋白海绵(fibrin collagen sponge)，B组：生物蛋白海绵(fibrin collagen sponge and bFGF);C组：明胶海绵(gelatin sponge)3种载体上，于4周取材进行组织学分析。[结果]纤维胶原蛋白海绵和生物蛋白海绵组均有软骨细胞形成，明胶海绵无细胞生长。利用SPSS 10.0软件统计学处理，显示B组软骨细胞阳性数量(92.75±10.57)多于A组(36.45±8.34)，有统计学差异(Plt;0.01)，两组都明显多于C组(0)，有统计学差异(Plt;0.01)。[结论]MSCs经分离扩增后种植于三维立体结构的纤维蛋白海绵载体上，在含有TGFβ1培养液中形成软骨;在TGFβ1和FGF的双重因子培养中，其软骨的形成能力明显增加。 【关键词】 骨髓基质细胞; 载体; 软骨形成; 组织工程 　　Abstract：[Objective]To investigate the chondrogenic ability and feasibility of bone mesenchymal stem cells (BMSCs) in three kinds of cellular carrier.[Method]BMSCs were separated and cultured into the third generation,which were cultivated in three knids of carriers by multipoint injection into and dropping on the surface of carriers(fibrin collagen sponge, group A; fibrin collagen sponge and FGF,group B and gelatin sponge, group C ).All samples were analyzed after four weeks.[Result]There were a few chondrogenic cells in group A,much more chondrogenic cells in group B,and no cells in group C. According to analysis with SPSS 10.0 software,There were more cartilage cells (92.75±10.57) in group B than in group A(36.45±8.34).It was statistical different (Plt;0.01).[Conclusion]BMSc can be formed to chondrogenesis after augmented and cultivated in the threedimensional fibrin collagen sponge with the medium of contained transforming growth factorβ1.Its ability of chondrogenesis is increased evidently in cultured by diplex growth factors(TGF and FGF). 　　Key words：bone mesenchymal stem cells; cellular carrier; chondrogenesis; tissue engineering 　　骨髓干系胞(bone marrow stromal cells，BMSCs)也称骨髓的间充质干细胞(bone mesenchymal stem cells，BMSCs)，BMSCs的提取分离及传代培养技术已逐渐成熟。如何将BMSCs种植于载体上进行体外人工培养形成软骨样组织还是在探讨中的问题。本研究旨在探讨BMSCs能否在体外几种载体中提供软骨诱导微环境，促进BMSCs向软骨分化并形成组织工程软骨。 　　1 材料与方法 　　1.1 细胞培养 　　BMSCs培养：抽取8周龄幼猪髂骨骨髓5 ml，用密度梯度离心法分离出有核细胞，再以PBS缓冲液(Gibco公司，美国)和无血清DMEM(Gibco公司，美国)各离心洗涤1次。制备含