兔骨髓间充质干细胞体外诱导软骨形成

目的：体外分离培养兔骨髓间充质干细胞(MSCs)，在诱导刺激物作用下，使其分化为软骨细胞。方法：在骨髓MSCs培养液内加入地塞米松10_-7 mol?L^-1，维生素C 50 mg?L^-1， 基因工程牛酸性成纤维细胞生长因子(bFGF)10 μg?L^-1，培养1周后将bFGF换为基因工程人转化生长因子-β₁(TGF-β₁)10 μg?L^-1，继续培养3周。 结果：细胞形态由纺锤形变为圆形或椭圆形，细胞分泌了大量的甲苯胺蓝异染的基质，形成了类软骨陷窝。碱性磷酸酶(ALP)的活性增高了近20倍。逆转录PCR证实转化前的细胞主要表达Ⅰ型胶原mRNA。转化后的细胞主要表达Ⅱ型胶原mRNA。 结论：在该种诱导环境下,部分骨髓MSCs可以转化为软骨细胞。

Chondrogenic induction of rabbit bone marrow mesenchymal stem cells in vitro

ObjectiveTo isolate and culture the rabbit bone marrow mesenchymal stem cells (MSCs) and then to induce MSCs into chondrocytes in vitro .MethodsThe MSCs was isolated and expanded from cultured and nucleated cell fraction of autologous bone marrow. Dexamethasone(10 -7 mol·L -1 ), ascorbic acid (50 mg·L -1 ) and basic fibroblast growth factor (bFGF 10 μg·L -1 ) were added into MSCs medium for the first week. Then the bFGF was replaced by transforming growth factor β 1 (TGF β 1 10 μg·L -1 ) for next three weeks. ResultsSome spindle MSCs turned into round or elliptic shape and secreted metachromatic matrix with Toluidine blue. The alkaline phosphates(ALP) activity of MSCs increased to twenty fold . The RT PCR indicated that the MSCs mainly expressed type Ⅰ collagen mRNA before induction. After induction the partial differentiated cells begun to express type Ⅱ collagen mRNA.ConclusionIn this condition the rabbit MSCs may be induced into chondrocytes in vitro .