Settmacher U, Jahn S, Grunow R, Mehl M, von Baehr R
Department of Medicine (Charité), Humboldt-University, Berlin.
Allerg Immunol (Leipz). 1989;35(3):195-201.
The aim of this study was to develop an optimal technique for cryopreservation of newly formed human and mouse hybridoma cells immediately after fusion. It was shown that human hybridoma cells could be frozen most successfully at a cooling rate of 5 K/min whereas mouse hybridomas at 1 K/min. The percentage of FCS in cryopreservation media (30 or 90%) had no influence on the recovery of both types of hybridomas. For testing the efficiency of freezing, the fusion rate (number of growing and Ig producing hybridomas), the ratio of IgM:IgG-producing initial hybridoma lines and yields of specific wells were analysed. Comparable results were registered when optimally cryopreserved and non-frozen material was studied. Optimization of the cryopreservation of newly formed hybridomas may be of methodological importance, especially when a time-consuming screening for antigenic specificities must be carried out.
本研究的目的是开发一种在融合后立即对新形成的人源和鼠源杂交瘤细胞进行冷冻保存的最佳技术。结果表明,人源杂交瘤细胞在5 K/分钟的降温速率下冷冻最为成功,而鼠源杂交瘤细胞在1 K/分钟的降温速率下冷冻效果最佳。冷冻保存培养基中胎牛血清(FCS)的百分比(30%或90%)对两种类型杂交瘤细胞的复苏均无影响。为测试冷冻效率,分析了融合率(生长且产生免疫球蛋白的杂交瘤细胞数量)、产生IgM与IgG的初始杂交瘤细胞系的比例以及特定孔的产量。对最佳冷冻保存和未冷冻的材料进行研究时,得到了可比的结果。新形成的杂交瘤细胞冷冻保存的优化可能具有方法学重要性,特别是在必须进行耗时的抗原特异性筛选时。