保护剂和冻存时间对昆虫细胞系NIAS-MaBr-85冻后活性的影响  

Effects of Cryoprotectants and Durations of Cryopreservation on Post-thaw Cytoactive of Insect Cell Line NIAS-MaBr-85

作　　者：马艳[1] 丁伟峰[1] 冯颖[1] 张欣[1] 马涛[1] 

机构地区：[1]中国林业科学研究院资源昆虫研究所国家林业局资源昆虫培育与利用重点实验室,昆明650224

出　　处：《中国细胞生物学学报》2010年第1期154-158,共5页Chinese Journal of Cell Biology

基　　金：林业公益性行业科研专项(4-38);国家林业局948项目(2002-52)资助~~

摘　　要：本文分别使用3种不同配方的冻存保护液对甘蓝夜蛾(Mamestra brassicae)5龄幼虫脂肪体细胞系NIAS-MaBr-85进行液氮低温冷冻保存,并在第1、3、9、17、20、28、38个月时检测细胞活力和圆度的变化情况,以评估不同冷冻保护剂和冻存时间对细胞活性的影响。研究发现,NIAS-MaBr-85细胞的冻后活性随着冻存时间的延长而降低。短期冻存时,3种冻存保护液对NIAS-MaBr-85的冻后活性影响不明显,在长期冻存条件下,以10%DMSO作为冻存保护剂的细胞冻后活力和圆度变化幅度明显小于以10%甘油为保护剂冻存的细胞。使用胎牛血清(FBS)替代培养基进行细胞冻存,对保持细胞冻后活性稳定的作用十分有限。因此,10%DMSO是长期冻存NIAS-MaBr-85细胞的优良冻存保护剂。通过对不同冻存时间的细胞活性比较发现,虽然冻存时间的长短对于细胞冻后活力和圆度变化有显著影响,但在昆虫细胞的长期低温保存过程中,冻存保护剂的差异是影响冻后细胞生物学活性的主要因素。In order to evaluate the effects of different cryoprotectants and durations on post-thaw cytoactive of insect cell line NIAS-MaBr-85, 3 kinds of cryoprotectants （Formula Ⅰ： 10% DMSO ＋ 90% FBS; Formula Ⅱ： 10% DMSO; Formula Ⅲ： 10% glycerol） were used to preserve cells in liquid nitrogen. The post-thaw viability and circularity of cells were examined at 1, 3, 9, 17, 20, 28, and 38 months separately. The results showed that the post-thaw viability of cells decreased with the time of cryopreservation expended. In short-term cryopreservation, the effects of different cryoprotectants on post-thaw cytoactive of NIAS-MaBr-85 were not significant. In long-term cryopreservation, the effect of preservation of 10% DMSO was better than that of 10% glycerol. The effect of supplement of 90% FBS on preserving the post-thaw biological characteristics of cells was not obvious. Comparing the influence of the length of cryopreserved time, different cryoprotectants had a statistically significant influence on the post-thaw biological characteristics of NIAS-MaBr-85 cells.

关 键 词：冷冻保护剂 冻存时间 昆虫细胞 细胞活力 

分 类 号：Q962[生物学—昆虫学]