不同冻存液冻存慢病毒转染后稳转肝癌细胞株HepG2的效果比较

The optimal cryopreservation liquid of the stable transfected cell line named hepatic carcinoma HepG2 Transfected by Lintivirus

目的筛选冻存慢病毒转染后稳转肝癌细胞株HepG2的最佳冻存液。方法采用甘油和二甲基亚砜(DMSO)2种不同冻存保护剂,以4种不同比例的冻存液分别冻存慢病毒转染后肝癌细胞HepG2及未转染的HepG2细胞,并对上述细胞复苏后的存活率及增殖能力进行比较分析。结果采用甘油为冻存保护剂时,冻存液[DMEM∶胎牛血清(FBS)∶甘油]比例为0∶9∶1的转染后细胞存活率最高(P=0.001);未转染细胞存活率与冻存液比例无关(P=0.293)。采用DMSO为冻存保护剂时,转染后细胞存活率为0%;未转染细胞存活率在60%及以上,且与冻存液比例无关(P=0.487)。冻存液中血清含量越高,复苏后细胞的增殖能力越强(P<0.05)。结论冻存慢病毒转染后的稳转肝癌细胞株HepG2的最佳冻存液为90%FBS+10%甘油。

Objective To investigate the optimal cryopreservation liquid of the stable transfeeted cell line named hepatic carcino- ma HepG2 transfected by lentivirus. Methods Glycerol and DMSO as cryopreservation liguid,The HepG2 transfected by lentivirus and HepG2 cells were both cryopreserved with four various proportion of cryopreservation liquid. After thawing, the survival rate of the two cells were observed by inverted contrast microscope and the viability and proliferation were detected with MTT assay. Re- sults Using glycerol as cryoprotectant liquid, the survival rate of the HepG2 cells transfected by lentivirus was obviously higher than other 3 groups when the proportion of the cryopreservation liquid（DMEM ： FBS ： Glycerol）was 0 ： 9 ： 1（P=0. 001） ,where- as there was no significant difference among the groups of HepG2 cells with various proportion of cryopreservation liquid（P = 0. 293）. Using DMSO as eryoprotectant liquid, the survival rate of the cells transfected by lentivirus was 0 %, whereas the survival rate of the HepG2 cells was higher than 60% ,and there was no significant difference between groups of various proportion of cryo- preservation liquid（P= 0. 487）. The MTT assay demonstrated that the higher the serum levels was, the better proliferation capacity those cells had（P〈0.05）. Conclusion The optimal cryopreservation liquid for the hepatic carcinoma cell line HepG2 transfected by lentivirus is used glycerol as cryoprotectant solution, and the proportion of cryopreservation solution was 90% FBS＋ 10% Glycerol.