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.

  • Day 1-Morphology

HUREL® Primate™

Phase contrast image in a 96-well at a 10× magnification. Image Credit: Visikol Inc.

  • Day 7-Morphology

HUREL® Primate™

Phase contrast image in a 96-well at a 10× magnification. Image Credit: Visikol Inc.

  • Day 7-Bile Canaliculi

HUREL® Primate™

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

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