外源性透明质酸对兔骨髓间充质干细胞定向分化为软骨细胞的影响

发布时间：2018-01-10 22:13

  本文关键词：外源性透明质酸对兔骨髓间充质干细胞定向分化为软骨细胞的影响　出处：《青岛大学》2011年硕士论文　论文类型：学位论文

  更多相关文章： 骨髓间充质干细胞 软骨细胞 透明质酸 分化

【摘要】：背景：骨髓间充质干细胞有多向分化潜能,在不同的诱导条件下可以分化成许多不同的组织,近年来已成为组织工程软骨构建过程中最常用的种子细胞。 目的：通过分析外源性透明质酸对兔骨髓间充质干细胞体外增殖及定向分化为软骨细胞的影响,探讨关节腔内环境对骨髓间充质干细胞的作用。 方法：全骨髓法+贴壁培养法分离培养兔骨髓间充质干细胞,取第4代细胞用于实验,实验组细胞加入透明质酸诱导液,以转化生长因子β3诱导组作为阳性对照,阴性对照组加入常规培养液。分别于诱导后第7,14,21 d行甲苯胺蓝染色检测蛋白聚糖表达,免疫组化染色及RT-PCR检测细胞Ⅱ型胶原表达。 结果：经透明质酸诱导后,细胞增殖速度减慢,由长梭形变为多角形、椭圆形,细胞外基质呈甲苯胺蓝异染性和Ⅱ型胶原免疫组化阳性, RT-PCR检测示Ⅱ型胶原mRNA表达阳性,表现出软骨细胞的分化特点,但表达均比阳性对照组弱。 结论：外源性透明质酸具有诱导兔骨髓间充质干细胞向软骨细胞分化的能力,但比转化生长因子β3的诱导能力弱。结果提示,关节腔内环境对骨髓间充质干细胞向软骨细胞分化有正性促进作用,支持透明质酸作为软骨组织工程基质使用。
[Abstract]:Background: bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into many different tissues under different induction conditions and have become the most commonly used seed cells in the process of tissue engineering cartilage construction in recent years. Aim: to investigate the effects of exogenous hyaluronic acid on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells into chondrocytes in vitro. Methods: the rabbit bone marrow mesenchymal stem cells were isolated and cultured by whole bone marrow adherent culture method. The cells of the 4th passage were used in the experiment. The cells in the experimental group were added hyaluronic acid inducer. Transforming growth factor 尾 3 group was used as positive control group and negative control group was added into conventional culture medium. Toluidine blue staining was used to detect proteoglycan expression on day 71421 after induction. The expression of type 鈪，

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