Pĕknicová J, Kristofová H
Folia Biol (Praha). 1985;31(5):357-60.
Stabile hybridoma cells, colonies of hybridoma cells 14 days after fusion of immune spleen and myeloma cells, myeloma cells and fibroblasts cultured in 96-well microculture plates were frozen by the method of two-step freezing. The culture medium was aspirated, and 50 microliter of the medium containing a cryoprotectant (5% dimethyl sulphoxide) was added for 10 min at room temperature. The plates were put into microtene bags, placed at -25 degrees C in a freezer for 30 min and then stored at -100 degrees C in liquid nitrogen vapour. Plates with cells were thawed rapidly in a 50 degree C water bath. After thawing the hybrid cells were viable and continued to produce the specific antibody.
稳定的杂交瘤细胞，即免疫脾细胞与骨髓瘤细胞融合14天后的杂交瘤细胞集落，以及在96孔微量培养板中培养的骨髓瘤细胞和成纤维细胞，采用两步冷冻法进行冷冻。吸出培养基，加入50微升含有冷冻保护剂（5%二甲基亚砜）的培养基，在室温下放置10分钟。将培养板放入微量袋中，置于-25℃的冰箱中30分钟，然后在液氮蒸汽中-100℃保存。装有细胞的培养板在50℃水浴中快速解冻。解冻后杂交细胞仍具有活力，并继续产生特异性抗体。
