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.

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.