Overview

A co-culture called HUREL® Human Pool™ is made up of a collection of primary human hepatocytes that have been cryopreserved and are then cultured with non-parenchymal stromal cells.

Example Metabolic Activity (nmoles/hr/106 cells). Source: Visikol Inc.

Substrate Enzyme Concentration (µM) Day 1 Day 4 Column 8
Midazolam CYP3A4 5 1.153 1.363 1.077
Dextromethorphan CYP2D6 20 0.50 0.440 0.9
Tolbutamide CYP2C9 20 0.044 0.091 0.231

 

Pooled cryopreserved human hepatocytes were defrosted, plated on HUREL PlatinumHeps™ Media supplemented with 10% serum, and then switched to HUREL PlatinumHeps™ Basal Media 24 hours after seeding.

The concentrations of CYP substrates are listed in the table above, 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.

  • Day 1-Morphology

HUREL® Human Pool™

Phase contrast image in a 24-well at a 10× magnification. Image Credit: Visikol Inc.

  • Day 7-Morphology

HUREL® Human Pool™

Phase contrast image in a 24-well at a 10× magnification. Image Credit: Visikol Inc.

  • Day 7-Bile Canaliculi

HUREL® Human Pool™

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 10× magnification with filters EX/EM 492-495/512-527 nm. Image Credit: Visikol Inc.

Example culture origin

Human donor demographics

  • Gender: Male (2) Female (3)
  • Age (years): 40–70
  • Race: Caucasian. (4) African American (1)
  • Cause of Death: Anoxia (1), N/A (4)
  • Tobacco History: Not (2) Past Use (3)
  • Alcohol History: Past Use (2) Current Use (3)
  • Medical History: Not Given

Donor serology

  • Hepatitis B: Negative
  • Hepatitis C: Negative
  • HIV: Negative
  • EBV: IgG [+] (5), IgG [-} (1), N/A (4)
  • RPR: Negative
  • CMV: Positive (6) Negative (4)

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