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The uptake properties of taurocholate (TC) and estradiol 17β-D-glucuronide (E₂17βG) were examined in freshly isolated and cryopreserved human hepatocytes to discover if active transport is retained in cryopreserved human hepatocytes. Firstly, the uptake of TC and E₂17βG was measured before and after cryopreservation. The uptake of TC was found to be Na⁺-dependent both in fresh and cryopreserved hepatocytes. The uptake activity in cryopreserved hepatocytes was found to range from 10 to 200% of that observed in freshly isolated cells. A kinetic analysis was performed to evaluate the transport activity of TC and E₂17βG and revealed that the Michaelis constant (K_m) for these compounds in cryopreserved human hepatocytes was 2–8 and 3–18 μM, respectively. This was within the range of K_m values previously found in human Na⁺-taurocholate cotransporting polypeptides (NTCP) and organic anion …