Overview
HUREL® Cat™ is a co-culture made up of non-parenchymal stromal cells and cryopreserved primary feline hepatocytes.
Primary feline hepatocytes that had been frozen were thawed, plated on HUREL PlatinumHeps™ Media supplemented with 10% serum, and then switched to HUREL PlatinumHeps™ Basal Media 24 hours later. The concentrations of CYP substrates are listed in the table below, and metabolite formation is expressed as nmoles/hr/106 cells.
On Days 1, 4, and 8 after cell delivery, all incubations were performed in triplicate and incubated for 60 minutes. The reactions happened in a humid incubator with 5% CO2 at 37 °C. Supernatants collected were kept at –20 °C until they underwent additional LC/MS/MS analysis.
Example Metabolic Activity (nmoles/hr/106 cells). Source: Visikol Inc.
| Substrate |
Enzyme |
Concentration (µM) |
Day 1 |
Day 4 |
Day 8 |
| 7-Ethoxycoumarin |
Phase I |
100 |
0.110 |
0.117 |
0.059 |
| 7-Hydroxycoumarin |
Phase II |
100 |
5.060 |
3.950 |
4.277 |
| 7-Hydroxycoumarin |
Phase III |
100 |
3.687 |
3.250 |
4.090 |
Phase contrast image in a 24-well at a 10× magnification. Image Credit: Visikol Inc.
Phase contrast image in a 24-well at a 10× magnification. Image Credit: Visikol Inc.
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. Image Credit: Visikol Inc.
Example culture origin
Animal donor demographics
- Strain: Feline
- Number of Donors: 1
- Age (years): –
- Sex: Male