title: Plateable cryopreserved human hepatocytes for the assessment of cytochrome P450 inducibility: experimental condition-related variables affecting their response to inducers.
abstract: 1.
  
  
  BACKGROUND
  The aim of the present study was to assess the stability of cryopreserved human hepatocytes over 5 years and to explore experimental condition-related variables such as seeding density, culture matrix and medium, start and duration of treatment that could potentially affect the quality of cultures and their response to cytochrome P450 (CYP) inducers. 2.
  
  
  RESULTS
  63/125 batches of cryopreserved human hepatocytes were plateable after thawing. Of those, 17 batches showed reproducible recovery, viability and plateability (less than 5% intra-batch variability) up to 5 years. When cultured in collagen home-coated 48-well plates at a seeding density allowing 70% confluence, cryopreserved human hepatocytes display activities equivalent to fresh counterparts. Their response to CYP inducers is maximal and equivalent to fresh counterpart for an incubation of 72 h starting at Day 2 or Day 3 after plating when cultured in modified Hepatocyte Maintenance Medium (HMM). The number of cryopreserved human hepatocytes can be further reduced by using a cocktail of CYP substrates for the assessment of their inducibility. 3.
  
  
  CONCLUSIONS
  Experimental condition-related variables, such as seeding density, culture matrix and medium, start and duration of treatment, affecting the response of plateable thawed cryopreserved human hepatocytes to cytochrome P450 inducers can be reduced by optimizing critical steps of the protocols.
authors.affiliation: KaLy-Cell, 20A rue du Général Leclerc, Plobsheim, France.
authors.forename: Eliane
authors.initials: E
authors.lastname: Alexandre
authors.forename: Audrey
authors.initials: A
authors.lastname: Baze
authors.forename: Céline
authors.initials: C
authors.lastname: Parmentier
authors.forename: Coraline
authors.initials: C
authors.lastname: Desbans
authors.forename: Dumrongsak
authors.initials: D
authors.lastname: Pekthong
authors.forename: Brigitte
authors.initials: B
authors.lastname: Gerin
authors.forename: Claudine
authors.initials: C
authors.lastname: Wack
authors.forename: Philippe
authors.initials: P
authors.lastname: Bachellier
authors.forename: Bruno
authors.initials: B
authors.lastname: Heyd
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Weber
authors.forename: Lysiane
authors.initials: L
authors.lastname: Richert
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis*; D004790:Enzyme Induction; D022781:Hepatocytes / Q000166:cytology / Q000201:enzymology*; D006801:Humans; D008858:Microscopy, Phase-Contrast; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D003577:Cytochrome P-450 Enzyme System