摘要
目的体外诱导犬骨髓基质干细胞(BMSCs)定向分化为软骨细胞,探讨体外诱导成软骨的方法和条件。方法自犬肋骨取骨髓2～3ml,体外行原代和传代培养扩增,顺序加入碱性成纤维细胞生长因子(bFGF)和转化生长因子β1(TGF-β1),以培养瓶内较高细胞浓度培养,诱导BMSCs分化为软骨细胞。甲苯胺蓝、阿新蓝染色检测软骨基质的分泌,免疫组织化学染色检测软骨特异性Ⅱ型胶原表达。结果诱导的软骨样细胞甲苯氨蓝异染性、阿新蓝染色阳性;Ⅱ型胶原免疫组织化学检测阳性。结论应用bFGF和TGF-β1体外可以诱导犬BMSCs分化为软骨细胞,诱导的软骨细胞可作为软骨组织工程较理想的种子细胞。
Objective To explore the methods of the differentiation from canine bone marrow stem cells (BMSCs) into chondrocytes in vitro and determine the factors involved in the differentiation process. Methods About 2 to 3ml BMSCs were aspirated from canine ribs, primarily cultured and subcultured in vitro. After bFGF and TGF-β1 were added into the culture medium sequentially, BMSCs were cultured and expanded in the medium until they reached the required number. BMSCs were induced to differentiate into chondrocytes at high cell density. Matrix of cartilage cells was detected by toludine blue and alcian blue staining, and cartilage specific collagen Ⅱ was detected by immunohistochemistry. Results The structure of cellular cartilage form BMSCs was uniformly positive of alcian blue staining and toludine blue staining. Immunohistochemical staining was positive for the passage cells, Collagen Ⅱ. Conclusion Application of bFGF and TGF-β1 may induce canine bone marrow stem cells into chondrocytes in vitro, which can be used as seeding cells in chondrocyte tissue engineering
出处
《中华创伤骨科杂志》
CAS
CSCD
2004年第7期763-766,共4页
Chinese Journal of Orthopaedic Trauma
基金
广东省自然科学基金
