微囊化軟骨細胞誘導骨髓間充質(zhì)干細胞的體外定向分化
發(fā)布時間：2018-08-27 08:31
【摘要】：背景:以往誘導骨髓間充質(zhì)干細胞定向分化的直接接觸共培養(yǎng)法、Transwell膜非接觸共培養(yǎng)誘導法存在干細胞純度低、增殖緩慢等不足。目的:對比分析微囊化共培養(yǎng)體系與傳統(tǒng)Transwell共培養(yǎng)體系對骨髓間充質(zhì)干細胞的誘導效果。方法:實驗組取第3代兔骨髓間充質(zhì)干細胞與微囊化的第2代兔關節(jié)軟骨細胞,按1∶1的比例在Transwell小室進行共培養(yǎng),同時設置第3代兔骨髓間充質(zhì)干細胞與第2代兔關節(jié)軟骨細胞共培養(yǎng)于Transwell小室的傳統(tǒng)共培養(yǎng)組,以及純干細胞培養(yǎng)組。MTT法對比3組干細胞的增殖率,甲苯胺藍染色、番紅花O染色觀察軟骨基質(zhì)的合成,阿利新藍染色法和Elisa法定量測量糖胺聚糖與Ⅱ型膠原蛋白的合成。結果與結論:傳統(tǒng)共培養(yǎng)組細胞增殖率低于實驗組與純干細胞培養(yǎng)組(P0.05)。實驗組細胞糖胺聚糖和Ⅱ型膠原蛋白水平高于傳統(tǒng)共培養(yǎng)組、純干細胞培養(yǎng)組(P0.05),甲苯胺藍染色、番紅花O染色強度強于傳統(tǒng)共培養(yǎng)組、純干細胞培養(yǎng)組。表明微囊化的軟骨細胞能夠成功誘導骨髓間充質(zhì)干細胞向成軟骨方向定向分化,且誘導效果優(yōu)于傳統(tǒng)的Transwell膜分離共培養(yǎng)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.
【作者單位】： 西安交通大學第二附屬醫(yī)院耳鼻咽喉頭頸外科病院;西安交通大學第二附屬醫(yī)院骨科;
【基金】：“中央高�；究蒲袠I(yè)務費專項資金”資助(0817-08143017)~~
【分類號】：R329
，
本文編號：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.
【作者單位】： 西安交通大學第二附屬醫(yī)院耳鼻咽喉頭頸外科病院;西安交通大學第二附屬醫(yī)院骨科;
【基金】：“中央高�；究蒲袠I(yè)務費專項資金”資助(0817-08143017)~~
【分類號】：R329
，
本文編號：2206689
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