自1977年以来,我们应用四级梯度降温冻存法对30余株哺乳动物细胞及2株节肢动物细胞共1000多支安瓿进行了冷冻保存,效果较好,大部分细胞的活存率在70％以上。在此基础上对8株抗登革4型(DEN-4)病毒杂交瘤细胞400多支安瓿进行了冻存。鉴于四级降温冻存法步骤多,需要经过40℃冰箱和一20℃冰箱长达7～8小时的梯度降温,较难适应单克隆抗体工作中细胞冻存的需要。因此,自1982年以来我们摸索了影响细胞冻存的主要因素,确定了杂交瘤细胞冻存的最适保护剂浓度,并研制成功细胞冻存简易装置。冻存操作时间缩短了4～5小时,减少 Since 1977, we have used four-step gradient cryopreservation method to cryopreserve more than 1000 ampoules of more than 30 mammalian cells and 2 arthropod cells, and the effect is good. Most of the cells have a viability of 70 %the above. On this basis, more than 400 anti-dengue 4 (DEN-4) virus hybridoma cells were cryopreserved over 400 ampoules. In view of the four steps of cryopreservation method more steps, need to go through a 40 ℃ refrigerator and a 20 ℃ refrigerator up to 7 to 8 hours of gradient cooling, more difficult to adapt to the work of monoclonal antibody cryogenic needs. Therefore, since 1982, we have explored the main factors that affect cell cryopreservation, determined the optimum concentration of cryoprotectants for hybridoma cells, and developed a simple cell cryopreservation device. Frozen storage operation time reduced by 4 to 5 hours, reduced