Procedure | CCHH | MMHH |
---|---|---|
Vial retrieval of from storage | From liquid nitrogen storage (3–5 minutes) | From −80°C storage (3–5 minutes) |
Thawing of the cryopreserved hepatocytes in a 37°C water bath | 1.5–2 minutes | 1.5–2 minutes |
Addition of the thawed hepatocytes to recovery medium followed by centrifugation for 10 minutes for cell recovery | 10–15 minutes | Not required |
Microscopic examination of thawed hepatocytes for the quantification of viability and cell concentration | 10–15 minutes | Not required |
Adjustment of cell suspension to contain 2× the final desired cell density | 5 minutes | Not required |
Addition to vessels containing 2× test compoundsa | ||
Total time from freezer to incubation | 30.5–45 minutes | 7–10 minutes |
↵a For example, addition of 50 μl of hepatocytes to a 96-well plate with 50 μl of test articles at 2× the final concentration, followed by incubation for the desired time duration at 37°C: 1–3 minutes.