Overview
Cryopreserved primary primate hepatocytes are co-cultured with cells of the stromal, non-parenchymal type in HUREL® Primate™.
Example Metabolic Activity (nmoles/hr/106 cells). Source: Visikol Inc.
| Substrate |
Enzyme |
Concentration (µM) |
Day 1 |
Day 4 |
Day 8 |
| 7-Ethoxycoumarin |
Phase I (ECOD) |
100 |
0.853 |
0.289 |
0.115 |
| 7-Hydoxycoumarin |
Phase II (UGT) |
100 |
78.33 |
71.5 |
84.5 |
| |
Phase II (SULT) |
– |
8.68 |
8.57 |
16.13 |
Hepatocytes from cynomolgus primate cryopreserved were thawed, plated on HUREL PlatinumHeps™ Media supplemented with 10% serum, and then switched to HUREL PlatinumHeps™ maintenance Media 24 hours after seeding.
The concentrations of CYP substrates are listed in the table above, and metabolite formation is expressed as nmoles/hr/million cells. On Days 1, 4, and 8 (the customer day), all incubations were performed in triplicate and incubated for 60 minutes.
The reactions happened in a humid incubator with 5% CO2 at 37 °C. Until a subsequent LC/MS/MS analysis, collected supernatants were kept at –20 °C.
Phase contrast image in a 96-well at a 10× magnification. Image Credit: Visikol Inc.
Phase contrast image in a 96-well at a 10× magnification. Image Credit: Visikol Inc.
Bile canaliculi assayed via 5-(and-6)-carboxy-2’, 7’-dichlorofluorescein diacetate (CDFDA) stain at a concentration of 5 µM and imaged in the GFP channel in a 96-well at 10× magnification. Image Credit: Visikol Inc.
Example culture origin
Animal donor demographics
- Strain: Cynomolgus
- Number of Donors: 3
- Age (years): 2~3
- Gender: Male