基于魔芋葡甘聚糖微球的胶原覆层型微载体的制备研究  

Preparation and Method of Microcarrier Based on Konjac Glucomannan Microsphere

作　　者：康跻耀[1,2] 张宁[1,2] 周炜清[2] 孙李靖[2] 张贵锋[2] 马光辉[2] 苏志国[2] 

机构地区：[1]北京中医药大学中药学院,北京100102 [2]中国科学院过程工程研究所生化工程国家重点实验室,北京100190

出　　处：《中国生物工程杂志》2013年第5期44-49,共6页China Biotechnology

基　　金：国家科技支撑计划项目(2012BAD32B09);国家"863"计划(2012AA021505);中科院知识创新项目(KSCX-EW-G-3)资助项目

摘　　要：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。Konjac glucomannan （KGM） microspheres were activated with 1,4-butanediol diglycidyl ether, and the collagen was coupled on the activated microsphere for preparation of micro-carrier for cell culture. For process optimization, the orthogonal regression experiments were carried out by four factors and three levels including activation time, protein consumption, coupled time, the crosslinker dosage effect of microcarrier cell culture. Based on culture effect of Vero cell, the optimum condition for preparation of collagen coated micro- carrier is activation time 5 h, the coupling of time 5 h, protein dosage 1 g： 0.1 g （ball： protein）, the amount of crosslinking agent lg： 0.5ml （bead： crosslinking agent）. The largest cell density was 1.7 × 106cells/ml under the optimal preparation condition. The result indicates that KGM microspheres coupled with collagen is suitable for cell culture as microcarrier.

关 键 词：魔芋葡甘聚糖微球 明胶 偶联 微载体 细胞培养 

分 类 号：Q78[生物学—分子生物学]