Each lot of human cryopreserved hepatocytes undergoes extensive quality checks including:

  • Morphology and cell health assessment
  • Metabolic activity testing
  • Genotyping analysis
  • Review of donor demographics
  • Application qualification

Morphology and cell health assessment (Figures 1 and 2)

  • Post-thaw viability ≥80% and stability
  • Cell shape and membrane integrity
  • Nucleus and organelle size and shape
  • Cytosolic clarity
  • Relative absence of cell debris
  • Cell–cell contacts and ≥80% confluency (plateable cells only)
  • Reestablishment of bile canalicular networks (plateable cells only)
  • Seeding density analysis

    Donor demographics

    • Gender
    • Age
    • Race
    • BMI
    • Serological data
    • Medications
    • History of smoking, alcohol, and drug abuse

    Metabolic activity tests

    • CYP1A2
    • CYP2B6
    • CYP2C8
    • CYP2C9
    • CYP2C19
    • CYP2D6
     
    • CYP2E1
    • CYP3A
    • FMO
    • ECOD
    • 7-HCG
    • 7-HCS
    • Others by request


    Application prequalification

    Human cryopreserved hepatocytes are tested and qualified for one or more of the following:

    • Suspension metabolism
    • Plated metabolism (intrinsic clearance)
    • CYP450 induction
    • Transporter uptake (suspension and plated)
    • Transporter uptake and efflux (plated)



    Figure 1. Comparison between poor and optimal human hepatocyte plated morphology. Hepatocytes isolated from separate donor tissues and cultured for several days produced divergent results - poor monolayer integrity was observed in lot A; whereas lot B exhibited optimal monolayer integrity.



    Figure 2. The relationship of seeding density to induction response. A recommended seeding density is supplied with each lot of GIBCO® human cryopreserved hepatocytes to ensure optimal induction response, as shown here with rifampicin.
    CMD SchemaApp code