Verification of the decrease in cell recovery after freezing and thawing due to suboptimal shipping using nine cancer cell lines and the differences in impacts between the cell lines

The impacts of suboptimal shipping conditions during transport on cell viability, recovery, and function of cryopreserved samples, have not been well studied.

The impacts of suboptimal shipping on viability and recovery after the freezing and thawing were investigated using nine cancer cell lines, with particular reference to the approximate level of exposure temperature and exposure time at which adverse effects occur, and whether there are differences in sensitivity between cell types.

The adverse effects of any set of suboptimal shipping conditions (-80 degree C for 7 d, -65 degree C or -50 degree C for 1, 3, and 7 d) on nine cancer cell lines (CHO-K1, COS-1, HeLa, HepG2, HL-60, Jurkat, MCF7, MDCK, 293T) were compared with data obtained during storage in liquid nitrogen.

No statistically significant decrease in viability was observed in seven of the nine cell lines after freezing and thawing. On the other hand, a statistically significant decrease in the cell recovery was observed after 2 d post freezing and thawing in the nine cell lines, except CHO-K1 at higher exposure temperatures and longer exposure times. Visualization of the adverse effects on the cell lines using a heat map showed that the impacts tended to be more pronounced under the condition of exposure at -50 degree C for three or more days.

These results will contribute to the development of standardized protocols and best practices for the optimal shipping of frozen animal cells. https://doi.org/10.54680/fr25210110212.