摘要
目的：观察冻存对分泌抗大肠癌SW1116细胞单克隆抗体（McAb）的杂交瘤细胞存活率的影响，以及对抗大肠癌SW1116细胞的McAb效价的影响。方法：杂交瘤细胞在12．5％DMSO下，于4℃、-30℃、-60℃及-196℃下分别冻存1、3、6、12及24个月，复苏，用台盼蓝拒染法测其存活率。McAb在等量甘油下，于4℃、-10℃、-20℃及-60℃下分别冻存1、3及12个月后，测其效价。结果：杂交瘤细胞置-196℃保存2年仍有78％的存活率；-60℃保存3、6及12个月的存活率分别为73％、40％及0％；不宜于-30℃及4℃保存。McAb在-20℃以下保存1年，效价未见下降；-10℃保存1年下降1个稀释度；4℃保存1年下降1—2个稀释度。结论：杂交瘤细胞短期保存可用-60℃冻存，长期需液氮保存。McAb长期保存最好于-20℃以下冻存。
Objective：To explore the optimal methods for cryopreservation of hybridoma cells, which can secrete monoclonal antibody （McAb） of anti- colorectal carcinoma cell line SW1116, and their secreting McAb. Methods： Hybridoma cells were cryopreserved in 12. 5% DMSO at 4℃, - 30℃, - 60℃, and - 196℃ for 1,3,6,12, and 24 months, respectively. Post - cryopreserved hybridoma cells were thawed and then their survival rates were determined by trypan blue staining test. Then the postthawed cells were observed. McAb was stored in 50% glycerol at 4℃, - 10℃, -20℃, and -60℃ for 1,3, and 12 months, respectively. Then the fiters of post - stored McAb were detected by ELISA. Results： When hybridoma cells were cryopreserved at - 196℃ for two years, their viability was 78%. When they were cryopreserved at -60℃ for 3,6,and 12 months,their viabilifies were 73% ,40% and 0% ,respectively. However, it was not suited to store the cells at 4℃ or -30℃. When McAb was stored at -20℃ or -60℃for one year, its titer was not decreased. When it was stored at - 10℃ or 4℃ for one year, its titer was decreased for 1 -2 dilutions. Condution：The hybridoma cells can be cryopreserved at - 196℃ for a long time, and also may be stored at -60℃ for a short period of time. The McAb can be stored at -20℃ or lower temperature for a long time.
出处
《中国卫生检验杂志》
CAS
2006年第5期536-537,共2页
Chinese Journal of Health Laboratory Technology
