Co-culture comprised of cryopreserved primary rat hepatocytes cultured with cells of non-parenchymal, stromal type.

Example Metabolic Activity (nmoles/hr/106 cells)

SubstrateEnzymeConcentration (µM)Day 1Day 4Day 8
7-EthoxycoumarinPhase I1000.0740.1830.181
7-HydroxycoumarinPhase II10014.3806.6408.460
7-HydroxycoumarinPhase II1003.8482.4841.572

Culture Conditions and Morphology

Cryopreserved primary rat hepatocytes were thawed and plated with HUREL PlatinumHeps™ Media supplemented with 10% serum and subsequently changed 24 hours post-seeding to HUREL PlatinumHeps™ maintenance Media. CYP substrate concentrations are given in the table above along with metabolite formation recorded as nmoles/hr/106 cells. All incubations were carried out in triplicate on days D1, D4, and D8 upon cell delivery and incubated for 60 minutes. Reactions took place in a humidified incubator at 37°C, in 5% CO2. Collected supernatants were stored at -20°C until further LC/MS/MS analysis.

Day 1 – Morphology

Day 7 – Morphology

Day 7 – Bile Canaliculi

Day 1- Rat Morphology phase contrast image

Phase contrast image in a 24- well at a 10x magnification

Day 7- Rat Morphology phase contrast image

Phase contrast image in a 24- well at a 10x magnification.

Day 7- Rat Bile Canaliculi assay

Bile canaliculi assayed via 5- (and-6)-carboxy-2’, 7’- dichlorofluorescein diacetate (C- DCFDA) stain at a concentration of 5 μM and imaged in the GFP channel in a 96-well at 10x magnification with filters EX/EM 492-495/512-527 nm.

Example Culture Origin

Rat Donor Demographics
  • Strain: Sprague-Dawley
  • Number of Donors: 16
  • Age: 2 months
  • Gender: Male