BMMSCs在温敏性玻璃微载体上的培养及收获

Culture and Harvest of BMMSCs on the Surface of Temperature-Responsive Glass Microcarriers

贴壁型种子细胞的培养和收获,常在二维静态的培养环境下进行,而后采用酶解法或机械刮刀法来进行收获,但是其存在着一定的弊端,细胞扩增数量有限且在收获时细胞易受到损伤,同时也会失去一定的干性。论文研究整合了具有较大比表面积和三维立体环境的微载体,及温敏性材料无损收获细胞的优点,采用表面自由基共聚合方法,用3-丙基三甲氧基硅烷甲基丙烯酸TMSPM为偶联剂,在微载体表面接枝上N-异丙基丙烯酰胺NIPAAm(N-isopropylacrylamide)及甲基丙烯酸羟基丙基酯(Hydroxypropyl methacrylate,HPM),制备出P(NIPAAm-co-HPM)-g-TMSPM-g-微载体。其次,将骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMMSCs)接种在制备的温敏性玻璃微载体上,通过细胞的培养与降温收获考察制备的温敏性微载体是否适宜细胞的降温脱附及其细胞相容性。扫描电镜及能谱分析表明温敏材料NIPAAm可成功地接枝到含硅羟基的玻璃微载体表面。BMMSCs的粘附和生长曲线测定结果证明材料具有较好的生物相容性,降温收获后细胞的活性对比及再培养实验表明细胞可很好的从载体上脱附,并保持良好的生长活性。

Attached seed cells are usually cultured in traditional two-dimension static plates and harvested via enzymatic or mechanical methods. However, these methods have disadvantages such as limited culture volume, cell damage and stemness losage. This study combined microcarriers（with large specific surface area and three-dimensional structure） and temperature-responsive materials to prepare P（NIPAAm-co-HPM）-gTMSPM-g-microcarrier, which was produced by grafting NIPAAm and HPM onto the surface of the microcarrier using TMSPM via surface free radical copolymerization. The performance of the temperature-responsive glass microcarrier was investigated by cell（Bone marrow mesenchymal stem cells, BMMSCs） culture and cooling harvest. SEM and EDS results show that NIPAAm can be successfully grafted and excellent biocompatibility is found from BMMSCs adhesion and growth curves. Most BMMSCs can detach from the microcarriers and keep viability.