Abstract: Konjac glucomannan microspheres were firstly aminated with 2-diethylaminoethyl chloride hydrochloride (DEAE), and activated with 1,4-butanediol diglycidyl ether. Then the charge-collagen double acting microcarrier was prepared via the coupling of the collagen on the activated microspheres. Furthermore, the effects of coating conditions, activation methods of collagen, sources of collagen and DEAE dosage on the microcarrier cell culture were examined. The results show that the collagen layer of charge-collagen double acting microcarrier can be controlled to 5~50 mm. After culture for 2 h, the cells attached to about 4×104 cells/mL, the attachment rate is between Cytodex 1 and Cydodex 3. The growth rate is faster than Cydodex 1 after 96 h, and cell number is more than Cytodex 1 and Cydodex 3 in final 144 h, reaching 1.59×106 cells/mL.

Key words: Konjac glucomannan microsphere, collagen, double acting microcarrier, cell culture

摘要： 以魔芋葡甘聚糖微球为基质，用2-二乙氨基氯乙烷盐酸盐(DEAE)将微球胺化，用1,4-丁二醇二缩水甘油醚活化微球，将胶原蛋白包覆到微球上，再用戊二醛交联，得到包覆均匀稳定的电荷-胶原蛋白双作用微载体. 考察了胶原蛋白包覆状况和活化方法、不同来源的胶原蛋白及DEAE用量对微载体细胞培养效果的影响，研究了Vero细胞在微载体上的培养情况. 结果表明，电荷-胶原蛋白双作用微载体的胶原层在5~50 mm范围内可控，培养2 h细胞的贴附量约为4×104 mL-1，介于Cytodex 1和Cydodex 3之间；96 h后微载体上细胞生长速度逐渐超过电荷型微载体Cydodex 1, 144 h时细胞数多于Cydodex 1和Cydodex 3，达1.59×106 mL-1.

关键词: 魔芋葡甘聚糖微球, 胶原蛋白, 双作用微载体, 细胞培养