兔骨髓间充质干细胞在藻酸锶凝胶中体外三维培养的实验研究Three- dimensional culture of rabbit bone marrow mesenchymal stem cells in Sr- alginate gels
涂以济;吴添龙;徐浚怀;叶爱芳;钟贞浩;程细高;
摘要(Abstract):
[目的]观察兔骨髓间充质干细胞(BMSCs)于藻酸锶凝胶中体外三维培养的形态与活性变化,为藻酸锶凝胶复合干细胞应用于骨组织工程提供理论依据。[方法]分离培养原代兔BMSCs并扩增,取第3代BMSCs以细胞密度为5×105个/ml复合于藻酸锶凝胶中三维培养,每日于倒置相差显微镜下观察细胞形态,在不同时间点对其进行Live/Dead染色测细胞存活,CCK-8检测细胞增殖,并用扫描电镜观察凝胶的超微结构及细胞于凝胶上的黏附情况。[结果]藻酸锶凝胶质软、透明、渗透性较好;BMSCs接种于藻酸锶凝胶中初始形态为圆形、呈悬浮生长,后细胞逐渐贴壁生长并呈星形或多角形;接种第1 d和第14 d凝胶中活细胞率分别为(88.61±4.08)%和(92.16±2.10)%,两者间无显著统计学差异(P>0.05);第4、7、10、13 d时二维培养组中OD值显著高于三维培养组(P<0.05);扫描电镜示藻酸锶凝胶呈典型的"鸡蛋箱"样结构,孔隙直径大小平均为150μm;接种于凝胶中7 d后可见细胞均匀黏附于凝胶壁生长。[结论]藻酸锶凝胶是一种优良的细胞支架,将其复合种子细胞应用于骨组织工程具有广阔的前景。
关键词(KeyWords): 骨髓间充质干细胞;藻酸锶;组织工程
基金项目(Foundation): 国家自然科学基金资助项目(编号:81060147)
作者(Author): 涂以济;吴添龙;徐浚怀;叶爱芳;钟贞浩;程细高;
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DOI:
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