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1 待克隆杂交瘤细胞的冻存杂交瘤技术中十分繁重费时的一个步骤是克隆化。如果要在一天内融合96孔8块板是比较困难的,一般要丢掉一部分。为了解决时间不足问题,作者采用了以下方法:先在显微镜下选好杂交瘤细胞,放入超净工作台,轻轻吸出上清液,加入3倍浓缩的细胞保护液,每孔2滴。培养板周围用酒精棉球擦净,封好。放入-10℃左右慢冻2h,然后放-70℃冰箱冻存。可在2～3天内随时取出进行挑选(3天后融合细胞死亡率升高)。用37℃温箱复苏或用常规方法均可。复苏后加入20％RPS营养液1滴,轻轻吸出1
1 A very laborious and time-consuming step in the cryopreservation hybridoma technique of hybridoma cells to be cloned is cloning. If it is difficult to fuse 8 plates of 96 wells in one day, it is generally necessary to lose part. In order to solve the problem of lack of time, the authors used the following method: first select the hybridoma cells under a microscope, place them in a clean bench, and gently suck the supernatant and add 3 times concentrated cell protection solution, 2 drops per well. Clean the plate around with alcohol cotton balls. Put in -10 °C or so slow-frozen 2h, then put -70 °C refrigerator frozen. Can be removed at any time within 2 to 3 days for selection (increased mortality of fused cells after 3 days). With 37 °C incubator or conventional method can be used. After resuscitation, add 1 drop of 20% RPS nutrient solution and gently aspirate 1