微囊化软骨细胞诱导骨髓间充质干细胞的体外定向分化
发布时间：2018-08-27 08:31
【摘要】：背景:以往诱导骨髓间充质干细胞定向分化的直接接触共培养法、Transwell膜非接触共培养诱导法存在干细胞纯度低、增殖缓慢等不足。目的:对比分析微囊化共培养体系与传统Transwell共培养体系对骨髓间充质干细胞的诱导效果。方法:实验组取第3代兔骨髓间充质干细胞与微囊化的第2代兔关节软骨细胞,按1∶1的比例在Transwell小室进行共培养,同时设置第3代兔骨髓间充质干细胞与第2代兔关节软骨细胞共培养于Transwell小室的传统共培养组,以及纯干细胞培养组。MTT法对比3组干细胞的增殖率,甲苯胺蓝染色、番红花O染色观察软骨基质的合成,阿利新蓝染色法和Elisa法定量测量糖胺聚糖与Ⅱ型胶原蛋白的合成。结果与结论:传统共培养组细胞增殖率低于实验组与纯干细胞培养组(P0.05)。实验组细胞糖胺聚糖和Ⅱ型胶原蛋白水平高于传统共培养组、纯干细胞培养组(P0.05),甲苯胺蓝染色、番红花O染色强度强于传统共培养组、纯干细胞培养组。表明微囊化的软骨细胞能够成功诱导骨髓间充质干细胞向成软骨方向定向分化,且诱导效果优于传统的Transwell膜分离共培养法。
[Abstract]:BACKGROUND: In the past, direct contact co-culture of bone marrow mesenchymal stem cells (BMSCs) induced by non-contact co-culture with Transwell membrane has some disadvantages, such as low purity and slow proliferation. The third generation of rabbit bone marrow mesenchymal stem cells and the second generation of rabbit articular chondrocytes were co-cultured in the Transwell chamber at a ratio of 1:1. Meanwhile, the traditional co-culture group of the third generation of rabbit bone marrow mesenchymal stem cells and the second generation of rabbit articular chondrocytes co-cultured in the Transwell chamber and the pure stem cell culture group were set up. Compared with the three groups, the proliferation rate of stem cells, toluidine blue staining, saffron O staining were used to observe the synthesis of cartilage matrix, Alixin blue staining and Elisa method to quantify the synthesis of glycosaminoglycan and type II collagen. Polysaccharide and type II collagen levels were higher than those of the traditional co-culture group, pure stem cell culture group (P 0.05), toluidine blue staining, saffron O staining intensity was stronger than that of the traditional co-culture group and pure stem cell culture group. The traditional Transwell membrane separation co culture method.
【作者单位】： 西安交通大学第二附属医院耳鼻咽喉头颈外科病院;西安交通大学第二附属医院骨科;
【基金】：“中央高校基本科研业务费专项资金”资助(0817-08143017)~~
【分类号】：R329
，
本文编号：2206689
[Abstract]:BACKGROUND: In the past, direct contact co-culture of bone marrow mesenchymal stem cells (BMSCs) induced by non-contact co-culture with Transwell membrane has some disadvantages, such as low purity and slow proliferation. The third generation of rabbit bone marrow mesenchymal stem cells and the second generation of rabbit articular chondrocytes were co-cultured in the Transwell chamber at a ratio of 1:1. Meanwhile, the traditional co-culture group of the third generation of rabbit bone marrow mesenchymal stem cells and the second generation of rabbit articular chondrocytes co-cultured in the Transwell chamber and the pure stem cell culture group were set up. Compared with the three groups, the proliferation rate of stem cells, toluidine blue staining, saffron O staining were used to observe the synthesis of cartilage matrix, Alixin blue staining and Elisa method to quantify the synthesis of glycosaminoglycan and type II collagen. Polysaccharide and type II collagen levels were higher than those of the traditional co-culture group, pure stem cell culture group (P 0.05), toluidine blue staining, saffron O staining intensity was stronger than that of the traditional co-culture group and pure stem cell culture group. The traditional Transwell membrane separation co culture method.
【作者单位】： 西安交通大学第二附属医院耳鼻咽喉头颈外科病院;西安交通大学第二附属医院骨科;
【基金】：“中央高校基本科研业务费专项资金”资助(0817-08143017)~~
【分类号】：R329
，
本文编号：2206689
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