HUREL® Human Pool™2021-10-14T12:37:20-05:00

HUREL® Human Pool™

Co-culture comprised of pooled cryopreserved primary human hepatocytes cultured with cells of non-parenchymal, stromal type.

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Example Metabolic Activity (nmoles/hr/106 cells)

SubstrateEnzymeConcentration (µM)Day 1Day 4Column 8
MidazolamCYP3A451.1531.3631.077
DextromethorphanCYP2D6200.50.440.9
TolbutamideCYP2C920.044.091.231

Culture Condition and Morphology

Pooled cryopreserved human hepatocytes were thawed and plated with HUREL PlatinumHeps™ Media supplemented with 10% serum and subsequently changed 24 hours post-seeding to HUREL PlatinumHeps™ Basal Media. CYP substrate concentrations are given in the table above along with metabolite formation recorded as nmoles/hr/106 cells. All incubations were carried out in triplicate on days 1, 4, and 8 upon cell delivery and incubated for 60 minutes. Reactions took place in a humidified incubator at 37°C, in 5% CO2. Col­lected supernatants were stored at -20°C until further LC/MS/MS analysis.

Day 1 – Morphology

day-1-human

Phase contrast image in a 24-well at a 10x magnification.

Day 7 – Morphology

day-7-morph

Phase contrast image in a 24-well at a 10x magnification.

Day 7 – Bile Canaliculi

day7-bile-1

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.

Example Culture Origin

Human Donor Demographics

  • Gender: Male(2) Female(3)
  • Age (yrs)” 40-70
  • Race: Cauc.(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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