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.