摘要
目的:以骨髓间充质干细胞作为种子细胞,探索组织工程化软骨的构建.方法:分离、获取、扩增骨髓间充质干细胞,通过将扩增的骨髓间充质干细胞接种于聚羟基乙酸(PGA)上,在成软骨诱导因子作用下在体外诱导培养3周后,将实验组和对照组的标本转植入自体兔腹腔内,进行体内培养6～12周后,再进行检测,评估工程化软骨的形成.结果:骨髓间充质干细胞接种于PGA支架上后,经过体外诱导培养和体内培养后,获得标本(7/10)出现软骨组织外观,进行组织学切片可见软骨陷窝,进行Ⅱ型胶原蛋白的免疫组化、Ⅱ型胶原mRNA原位杂交均为阳性.结论:兔骨髓间充质干细胞在成软骨诱导剂作用下,经体外和体内培养后,可生成组织工程化类软骨.
Objective： To explore the feasibility of constructing tissue- engineered cartilage by inducing rabbit bone marrow- derived mesenchymal stem cells（MSCs）. Methods： By Percoll density gradient centrifuge in vitro MSCs were isolated from rabbit bone marrow and were then purified in monolayer culture. MSCs were seeded onto polyglycolic acid（PGA） scaffolds in experimental groups to form cells - PGA composites which were subsequently cultured in a chemically defined medium mainly including transforming growth factor - （1 （TGF - β1, 10ng/ml）, dexamethasone（ 10^-7 mmol/ L）, ascorbic acid（50μg/ml）, 1%insulin, 1%selenium and 1%transferrin for three weeks in vitro. After that, cells- PGA composites in experimental groups were covered by greater omentum and implanted into the belly of adult New Zealan white rabbits for 6 - 12 weeks. At the same time, PGA scaffolds without MSCs in control groups were implanted in the same measure. Finally, morphological observation, hematoxylin-eosin（HE） staining, immunohistochemistry test and hybridization test in situ were conducted to assess cartilage regeneration after 6- 12 weeks. Results： MSCs were harvested from rabbit bone marrow and evenly attached to the surface of porous PGA scaffolds in experimental groups. Tissue - engineered specimens presented to be similar to cartilage in gross shape were successfully harvested in experimental groups, what＇s more, their specific markers were positive in the appearance of toluidine blue metachromasia and immunohistochemistry of type Ⅱ collagen and in situ hybridization of collagen type Ⅱ mRNA. Conclusion： Tissue - engineered cartilage could be harvested by inducing MSCs - PGA composites.
出处
《泸州医学院学报》
2005年第5期394-396,共3页
Journal of Luzhou Medical College
基金
四川省卫生厅科研资助资金(No.020060)
关键词
骨髓
干细胞
软骨.
Bone marrow
Stem cell
Cartilage
