HUREL® Human Pool™

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.

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

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

• Day 7-Morphology

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

• Day 7-Bile Canaliculi

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)