不同冻存条件下HepG_2细胞存活率的比较  

Comparing the effects of different conditions of cryopreservation on HepG_2 cell recovery rate

作　　者：苏秀珍[1] 朱晓莹[2] 姜艳[2] 曾怡[2] 

机构地区：[1]右江民族医学院广西少数民族体质特征研究实验室,广西百色533000 [2]右江民族医学院科学实验中心,广西百色533000

出　　处：《右江民族医学院学报》2016年第3期241-244,共4页Journal of Youjiang Medical University for Nationalities

基　　金：2014年广西高校科学技术研究项目(YB2014298);2014年右江民族医学院校级课题[右医科字(2014)2号]

摘　　要：目的探讨不同冻存条件下对HepG_2细胞存活率的影响,以提高复苏细胞的存活率。方法取对数生长期HepG_2细胞,分别配制传统冻存液A、改良冻存液B,采用方法1(人工梯度降温法)和方法2(程序降温盒法)两种冻存方法进行冻存,在冻存1个月、6个月、12个月后分别对细胞进行复苏,检测细胞的存活率并观察复苏细胞的生长状态。结果冻存1个月后复苏,冻存液A、B两组细胞的存活率差异无统计学意义(P>0.05),冻存6个月、12个月后复苏,冻存液A组细胞存活率显著低于B组(P<0.01);B组细胞复苏的生长状态及增殖速度都明显优于A组,两组细胞的数量差异有统计学意义(P<0.05);以方法1、2冻存的细胞,在冻存1个月、6个月、12个月后复苏细胞,方法2细胞的存活率均显著高于方法1(P<0.01)。结论采用含高浓度血清的冻存液及缓慢降温有利于提高细胞复苏的存活率。Objective To explore the influence of different cryopreservation conditions on HepG2 cell re-covery rate and to improve the resuscitation cell survival. Methods Logarithmic growth phase HepG2 cells were cryopreserved by using two different cell freezing medium （conventional freezing medium A and modified freezing medium B were prepared,respectively）and by using two methods （Method 1 ,artificial gradient cool-ing method;Method 2,programmed cooling box）.Cells were resuscitated at 1 month,6 months and 12 months after cryopreservation,respectively.Then the cell recovery rate was measured and cells growth status was observed by microscope. Results The resuscitation cell recovery rate was compared between freezing medium A group and freezing medium B group at 1 month after cryopreservation and there was no statistical difference （P〉0.05）.The cells were resuscitated at 6 and 12 months after cryopreservation,freezing medium A group had lower cell recovery rate than freezing medium B group comparison between the two groups showed statistically significant difference （P〈0.01）;and the resuscitated cells growth status and proliferated speed in group B were superior to group A,comparing the cell count between the two groups showed statistical differ-ence （P〈0.05）.The cells cryopreserved by Method 1 and Method 2 were resuscitated at 1 month,6 months and 1 2 months after cryopreservation.Method 2 had all higher cells recovery rate than Method 1 ,comparison the recovery rate between the two methods showed statistically significant difference （P〈0.01）. Conclusion The resuscitated cell recovery rate will be raised with a higher serum levels and programmed controlled cool-ing.

关 键 词：细胞培养技术 冷冻保存 存活率 

分 类 号：R394.26[医药卫生—医学遗传学]