Retention of transporter activities in cryopreserved, isolated rat hepatocytes

摘要: The success of cryopreservation isolated hepatocytes with existing methodologies is assessed respect to the retentivity cell integrity/viability (defined by trypan blue) and metabolic activities upon thawing in comparison those freshly prepared cells. But ability cryopreserved cells transport xenobiotics relative that has not been investigated. In this study, we optimized our previous methodology for evaluated metabolism thawed hepatocytes. Half freshly, rat collagenase perfusion were immediately used studies [14C]taurocholate, [3H]estrone sulfate [3H]estradiol 17β-d-glucuronide (1 μM) 7-hydroxy-4-(trifluoromethyl)-coumarin (100 μM), (3,4-difluorobenzyloxy)-5,5-dimethyl-4-(4-methylsulfonylphenyl)-(5 H )-furan-2-one (250 bufuralol tolbutamide probes UDP-glucuronyl transferase (UGT) CYP3A, CYP2D, CYP2C, respectively. remaining half was using an optimized, programmed-freezing protocol, which developed minimize prolonged release latent heat during freezing. With exception UGT probe, no significant difference ( P > 0.05) found both versus thawing. conclusion, have demonstrated first time a protocol retain drug activities.