牛肾细胞的冻存及复苏后连续传代的研究

Frozen storage and continuous passage after recovery of the bovine kidney cells

目的:研究牛肾细胞的冻存及复苏后连续传代的可行性,为轮状疫苗生产提供稳定、一致的细胞基质。方法:将原代牛肾细胞培养至一代消化后冻存,复苏后进行连续传代培养,观察和记录细胞生长情况。从原代开始,每隔5代对牛肾细胞做染色体核型分析,检查牛肾细胞传代稳定性。结果:牛肾细胞冻存后复苏可连续传至15代,细胞增殖数量、培养时间相对稳定,细胞生长良好,细胞形态未发生明显改变;牛肾细胞在传代过程中遗传性状稳定,未发现有染色体缺失或异倍体现象。结论:牛肾细胞冻存复苏后可连续传代培养,能够为轮状病毒疫苗生产提供稳定、一致的细胞基质。

Objective： To investigate the frozen storage and the feasibility of continuous passage after re- covery of bovine kidney cells, and to provide stable and homogeneous cell medium for the production of rotavirus. Methods： The primary bovine kidney cells were prepared and cultured to the first generation. Then the passage 1 cells were stored by cryopreservation after digestion. After recovery the cells was cultured and observed continuous- ly. From the primary passage, the number of chromosome was checked every 5 passages, and the passage stability of the bovine kidney cells was analyzed. Results： The bovine kidney cells were cultured for 15 passages after re- covery, the number of cell proliferation and culture time were stable, and the cell growth was well. Furthermore, none of cell morphological changes was observed. The chromosome numbers were stable, and no chromosome dele- tion or aneuploid occurred. Conclusion： The bovine kidney cells can be cultured continuously after frozen storage and recovery, thus they can provide stable and homogeneous cell medium for the production of rotavirus.