HUREL® Human™

Overview

Cryopreserved primary human hepatocytes are co-cultured with non-parenchymal stromal cells in HUREL^® Human™.

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

Human 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 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 microns 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
• Age (years): 65
• Race: Caucasian
• Cause of death: IHC-stroke
• Drug history: Not given
• Medications: Unknown statin, Lipitor, Lisinopril, Clonidine, and multivitamin
• Medical History: Basel cell: 15–20 years ago. Removed with no radiation-received no chemo or radiation with no reoccurrence. Hypertension diagnosed 20–25 years ago

Donor serology

• Hepatitis B: Negative
• Hepatitis C: Negative
• HIV: Negative
• EBV: IgG [+], IgM [–}
• RPR: Negative
• CMV: Positive