二甲基亚砜直接加入培养中Jurkat细胞进行冻存  

Dimethyl sulfoxide added into growing jurkat cells directly for freezing

作　　者：陈丽洁[1] 王犇[1] 赵起超[1] 马冠英[1] 童晨壕 陈功星[1] 

机构地区：[1]杭州师范大学医学院实验中心,浙江杭州310016

出　　处：《华夏医学》2016年第3期10-13,共4页Acta Medicinae Sinica

基　　金：浙江省科技厅分析测试科技计划资助项目(2015C37063);杭州师范大学本科生课外科研资助项目(2015R423052;CX2015127)

摘　　要：目的:探讨培养中的细胞直接加二甲基亚砜(DMSO)冻存的方法。方法:常规培养Jurkat细胞,接种换液培养过夜,然后封闭与未封闭室温继续培养,6 d后镜下观察生长状态;另外部分收集离心常规冻存,部分细胞直接加DMSO冻存,半年后复苏检测存活率和换液MTT分析细胞活力。结果:室温培养的Jurkat细胞,封闭组呈弱酸性,大部分细胞维持完整形态,少部分崩解,而未封闭组相反;常规和直接加DMSO组复苏存活率均超过50%,复苏过程中细胞活力逐渐上升,8 d后复苏成功可以传代,各检测点两组间均没有统计学差异(P>0.05)。结论:在培养中的Jurkat细胞直接加DMSO冻存,是一种新的细胞保存方法,可以作为细胞培养技术的一个补充。Objective： To investigate the method of directly adding dimethyl sulfoxide （DMSO） into growing jurkat cells for freezing. Methods： Jurkat cells growing normally were inoculated, and exchanged for cell medium and cultured overnight, then were divided them were cultured in room temperature. After into two groups： closed and non-closed sets. All of six days, they were observed and pictured under microscopy. In another experiment the overnight cultured cells were divided into two parts. One was collected by centrifuge and frozen regularly, the other was added DMSO into the Jurkat culture system directly. Then all of them were frozen in the liquid nitrogen. After 6 months, and the survival rate was checked with trypan blue at the time of recovery, and the cell viability was analyzed with MTT.Results： Under the room temperature, the closed set was with weak acid and the Jurkat ceils were much better and little broken in the weak acid, but the unclosed set was on the contrary. Furthermore, in the recovery time, the Jurkat survival rate was over 50% in both the convention and DMSO adding directly groups. Thecell viability was gradually increased in the process of recovery. After 8 days, the cell recovery was successful and subcuhured. There was no statistical differences between the convention and direct DMSO adding group in the all check points（P 〉 0.05）. Conclusion： The method of DMSO adding growing Jurkat cells is a new way of cryopreservation, which can be used as a supplement to the cell culture technique.

关 键 词：二甲基亚砜 细胞培养 冻存 

分 类 号：Q343.6[生物学—遗传学]