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以魔芋葡甘聚糖(KGM)微球为基质,用1,4-丁二醇二缩水甘油醚将KGM微球进行活化,将胶原覆层到微球上,对胶原覆层进行再次交联,得到覆层均匀、稳定的微载体。通过四因素三水平的正交回归组合试验设计,考察了活化时间、蛋白质用量、偶联时间、交联剂用量对微载体细胞培养效果的影响。以Vero细胞培养效果为指标,制备胶原包被微载体的最佳工艺为活化时间5 h、蛋白用量1∶0.1(球∶蛋白)、偶联时间5 h、交联剂用量每1gKGM加入0.5ml交联剂。在最优制备条件下,培养Vero细胞最大细胞密度可达到1.7×106cells/ml,证明了胶原覆层的KGM微球作为动物细胞培养的微载体具有可行性。
Using KGM microspheres as substrate, KGM microspheres were activated with 1,4-butanediol diglycidyl ether, collagen was coated on the microspheres and the collagen coating was re-crosslinked , Get coated uniform, stable microcarriers. Through four factors and three levels of orthogonal regression combined experimental design, the effects of activation time, protein dosage, coupling time and the amount of cross-linking agent on microcarrier cell culture were investigated. Vero cell culture effect as an indicator, the best preparation of collagen coated microcarriers activation time of 5 h, the amount of protein 1: 0.1 (ball: protein), coupling time 5 h, the amount of cross-linking agent per 1g KGM added 0.5ml Cross-linking agent. Under optimal conditions, the maximum cell density of cultured Vero cells reached 1.7 × 106cells / ml, demonstrating that collagen-coated KGM microspheres were feasible as microcarriers for animal cell culture.