CYP Induction in Cryopreserved Human Hepatocytes
Assay System:
- Plated cryopreserved human hepatocytes from 3 individual donors
- Induction of CYP1A2, CYP2B6 and CYP3A4 by test compound measured by comparing enzyme activity of hepatocytes treated with test compound with vehicle control
- Optionally, test compound concentration can be monitored during treatment in accordance with 2017 FDA guidance “In Vitro Metabolism- and Transporter-Mediated Drug-Drug Interaction Studies”
Assay Conditions:
| Test Compound Concentration | 30 µM, 3 µM, and 0.3 µM |
| Percent Organic Solvent – Inducer Treatment | 0.1% DMSO |
| Percent Organic Solvent – Substrate Incubation | 1.5% acetonitrile |
| Inducer Treatment Time | 48 hours |
| Substrate Incubation Time | 30 minutes |
| Treatment and Incubation Conditions | 37 °C, 5% CO2, and saturating humidity |
| Positive Control | Omeprazole (1A2), Phenobarbital (2B6), and Rifampicin (3A4) |
| Analysis | LC-MS/MS |
| Number of Repeats | 3 |
Deliverable: Mean fold induction of test compound and positive control compound.
Test Compound Requirement: 2-3 mg dry powder or 100-200 µL of 30 mM DMSO stock solution.