中国仓鼠卵巢细胞集落的简易原位贴壁冻存方法  

A simple method for in situ cryopreservation of CHO colonies

作　　者：卢柏松[1] 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所

出　　处：《军事医学科学院院刊》1998年第1期58-60,80,共4页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的：目前通用的细胞冻存方法是将细胞在保护剂的作用下悬浮冻存，这样细胞自然相互混合，复苏后不能保持原来的位置关系。本工作探索在需要时将中国仓鼠卵巢细胞（ＣＨＯ）在贴壁状态下原位冻存的可能性。方法：将转染促红细胞生成素（ＥＰＯ）表达质粒后形成的ＣＨＯ集落细胞在贴壁状态下加上１０％的二甲亚砜保护剂，然后分阶段降温并保存在－７０℃冰箱，两个月后复苏检测细胞的存活情况及其目的产物的表达情况。结果：集落至少可以保存两个月而绝大多数存活，而且从中能够得到表达外源目的基因的克隆。结论：将外源基因转染ＣＨＯ细胞后形成的克隆可以采用这种原位贴壁的方法冻存，这种冻存细胞的方法在需要保持细胞原来的位置关系时显然有用。Objectives: The routine method for the cryopreservation of CHO cells is to cryopreserve the cells in suspension, in which the cells are mixed and no longer maintain the original locations of the cells. The possibility of cryopreserving CHO cells in situ and adherently was studied. Methods: The CHO colonies formed after the transfection of an EPO activity expression plasmid into the CHO cells were cryopreserved adherently with the protectant 10% DMSO at -70℃. After two and a half months of cryopreservation the colonies were thawed and the survive rate and expression of foreign gene were assayed. Results: Over 90% of the colonies survived the cryopreservation and clones that express foreign gene could be obtained. Conclusion: It is possible to cryopreserve CHO colonies in situ and adherently. This simple method is especially useful when maintaining the original locations of the cells is essential in a study.

关 键 词：CHO细胞 细胞存活 贴壁低温冻存 低温保存 

分 类 号：Q2-33[生物学—细胞生物学]