HUREL® Rat WH™2021-10-14T12:38:38-05:00

HUREL® Rat WH™

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

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Example Metabolic Activity (nmoles/hr/106 cells)

Substrate

Enzyme

Concentration (µM)

Day 1

Day 4

Day 8

7-EthoxycoumarinPhase I(ECOD)C1000.050.050.05
Phase II(UGT)0.050.050.05
Phase II(SULT)0.050.050.05
7-HydroxycoumarinPhase II(UGT)1000.050.050.05
Phase II(SULT)0.050.050.05

Culture Conditions and Morphology

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

Morphology-Human-1-2

Phase contrast image in a 24-well at a 10X magnification

Day 7 – Morphology

Morphology-Human-2-2

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

Day 7 – Bile Canaliculi

Morphology-Human-3-2

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 10X magnifcation with filters EX/EM 492-495/512-527 nm.

Example Culture Origin

Human Donor Demographics

  • Gender: Male
  • Age (yrs)” 65
  • Race: Caucasian
  • Cause of Death: IHC-Stroke
  • Drug History: Not given
  • Medications: Unknown statin, Lipitor, Lisinopril, Clonaidine, 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
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