Cryopreservation of CHO Cells Using Serum-Free Media

초록

During routine maintenance, animal cell lines are commonly cryopreserved in growth medium containing with 10% DMSO. But,in case of bioprocess under the serum-free condition, including cultivation of cell lines and producing of pharmaceuticals,the cryopreservation should be executed without serum to prevent a cross-contamination. This experiments were performed to investigate the effects of the cryopreserved CHO cells in serum-free condition,first,the effects of permeable and non-permeable additives for suvstituge serum on cell viability were investigated. The combination of 10% DMSO and 0.3M raffinose in MEM-a without serum indicated 76% of cell viability. However, it did not reach the survival rates of the conventional cryopreservation. In the second, to evalutate the cryopreserved in the SFMs and the MEM-a with serum. All solution contained 10% DMSO.As a result of the comparison, cryppreserved CHO cells were thawed every three months to estimate the cell viability and the recovery rates. Then,real-time RT-PCR analyzed the inserted CHO DHFR gene. All result for the LTC appeared the same stability as the serum containing cryopreservation. In the conclusion,it could be seen that the LTC in the SFM can subsritute for serum using methods in the bioproceeed by CHO cells for more than two years.