大鼠多能成体祖细胞体外成软骨的诱导和鉴定  

Chondrogenesis of Rats' Bone Marrow-derived Multipotent Adult Progenitor Cells in vitro and Related Identification

作　　者：徐海艇[1] 潘骏[1] 李永领[1] 朱烈烈[1] 

机构地区：[1]温州医科大学附属第二医院育英儿童医院,浙江温州325027

出　　处：《浙江中医药大学学报》2016年第3期159-163,共5页Journal of Zhejiang Chinese Medical University

基　　金：浙江省自然科学基金资助项目(Y2100899);浙江省温州市科技局基金资助项目(Y20100026)~~

摘　　要：[目的]探讨大鼠多能成体祖细胞(multipotent adult progenitor cells,MAPCs)在特定培养条件下向软骨细胞定向分化的能力。[方法]分离培养大鼠MAPCs,扩增至第2代,观察其形态学特点,并将其离心成团后分为2组,诱导组应用转化生长因子-β3(transforming growth factor-β3,TGF-β3)、骨形态发生蛋白-6(bone morphogenetic protein-6,BMP-6)、胰岛素样生长因子-1(insulin-like growth factors-1,IGF-1)及地塞米松等进行成软骨诱导分化,对照组用含10%胎牛血清(fetal bovine serum,FBS)的高糖DMEM培养液(dulbecco's modifiedeagle medium high-glucose,H-DMEM)常规培养液自然分化。以Safranin O和Ⅱ型胶原免疫组化染色等进行鉴定,阿尔新蓝比色法测定糖胺聚糖(glycosaminoglycan,GAG)的含量。[结果]体外定向诱导后MAPCs表现出软骨细胞的特性。诱导组形成了透明软骨样外观的细胞团,而对照组体积小、松散,Ⅱ型胶原免疫组化染色显示诱导组可见成熟软骨陷窝形成,Safranin O染色显示诱导组软骨特异性细胞外基质沉积,诱导组GAG的含量高于对照组,差异有统计学意义(P<0.05)。[结论]MAPCs具有良好的成软骨潜能,可以成为软骨组织工程良好的种子细胞来源。[Objective] To explore chondrogenic differentiation of rats bone marrow-derived multipotent adult progenitor cells（MAPCs） in experimental culture medium.[Methods] MAPCs were isolated from rats bone marrow and expanded to passage 2. The morphology of MAPCs was observed in vitro. Then the cell groups were cultured in chondrogenic media of H-DMEM containing TGF-β3, BMP-6, IGF-1, the control one was made natural differentiation with routine fluid 10% FB5, and dexamethasone forming cell pellet and assessed by Safranin O and collagen Ⅱ staining. And Alcian blue chromatomery of glycosaminoglycan（GAG） was used to determine the differentiation. [Results] MAPCs demonstrated fibroblast-like morphology and showed good proliferation ability. After chondrogenic induction for 4 weeks, the pellets formed by MAPCs had a transparent white cartilage-like morphology in chondrogenic medium while a less shape in control medium. Histologically, the group showed a homogeneous cartilage-like structure with strong safranin O staining and strong collagen 2 staining. Alcian blue staining showed the content of GAG formatting were higher than the control group（P〈0.05）, with difference of statistical meaning. [Conclusion]MAPCs have high chondrogenic potential and could be an ideal seed in cartilage tissue engineering.

关 键 词：MAPCs 软骨细胞 诱导分化 软骨组织工程 

分 类 号：R331[医药卫生—人体生理学]