Co-culture comprised of cryopreserved primary rat hepatocytes cultured with cells of non-parenchymal, stromal type.
Example Metabolic Activity (nmoles/hr/106 cells)
|Substrate||Enzyme||Concentration (µM)||Day 1||Day 4||Day 8|
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
Phase contrast image in a 24- well at a 10x magnification
Phase contrast image in a 24- well at a 10x magnification.
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