摘要
目的 :体外定向诱导成人骨髓间充质干细胞 (MSC)分化为单一的软骨细胞 ,探索体外成软骨的必要条件。方法 :采用淋巴细胞分离液密度梯度离心法分离骨髓 ,体外培养扩增 ,在较高的细胞密度下加入转化生长因子 β1(TGF - β1) ,以微团培养 (micromassculture)方式 ,诱导MSC分化为软骨细胞。HE染色观察细胞形态 ,阿新蓝、甲苯氨蓝染色检测软骨基质的分泌 ,免疫组织化学检测软骨特异性Ⅱ型胶原表达。结果 :HE染色呈典型的细胞性软骨结构 ;阿新蓝染色阳性、甲苯氨蓝异染性 ;Ⅱ型胶原免疫组织化学检测阳性。结论 :成人骨髓MSC在体外分化为单一的软骨细胞需要高细胞密度、诱导因子TGF -
AIM: To investigate the differentiation of human bone marrow mesenchymal stem cells (MSC) into chondrocytes in vitro and determine factors involving in the differentiation process. METHODS: MSC were separated from iliac bone marrow with lymphocyte separating medium using density centrifugation. Cells were cultured and expanded in medium until reaching required number. MSC was induced to differentiate into chondrocytes by adopting high cell density, supplying growth factor and using micromass culture. Cells were observed by HE staining. Matrix of cartilage was detected by alcian blue and toludine blue and cartilage specific collagen II was detected by immunohistochemistry. RESULTS: The structure of the micromass assumed that of cellular cartilage, alcian blue staining were uniformly positive and toludine blue detected diffuse metachromasia substance, cells uniformly expressed collagen Ⅱ. CONCLUSION: High cell density, growth factor and appropriate culture conditions are critical to induce differentiation of MSC into chondrocytes. [
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第7期1195-1199,共5页
Chinese Journal of Pathophysiology
关键词
干细胞
软骨细胞
组织工程
Stem cells
Chondrocytes
Tissue engineering
