title: Morphological and biochemical integrity of human liver slices in long-term culture: effects of oxygen tension.
abstract: We tested the effects of low (20% O2) and high (70% O2) oxygen tension on the morphological and biochemical integrity of human liver slices incubated for up to 72 h in supplemented Williams' E medium in a dynamic rotating culture system. High oxygen tension was more effective than low oxygen tension for preserving morphological integrity in long-term culture (48-72 h). After 72 h of culture with 70% O2, the lobular pattern was well preserved, and the survival of hepatocytes (approximately 80%) and other cell types was good. Immunohistochemical studies showed good preservation of the region-specific expression of CYP2EI and CYP3A4 isoenzymes for up to 72 h of incubation in 70% O2. As compared to 20% O2, the oxidized glutathione content and reactive oxygen species production were slightly increased in 70% O2, suggesting that minimal oxidative stress occurred with the high oxygen tension. In conclusion, despite slight oxidative stress associated with high oxygen tension, 70% O2 appeared more appropriate than 20% O2 for preserving the morphological and biochemical integrity of human liver slices cultured in a dynamic organ culture system for up to 72 h.
authors.affiliation: Drug Safety Evaluation, Aventis Pharma SA, Vitry-sur-Seine, France.
authors.forename: H
authors.initials: H
authors.lastname: Martin
authors.forename: J P
authors.initials: JP
authors.lastname: Sarsat
authors.forename: C
authors.initials: C
authors.lastname: Lerche-Langrand
authors.forename: C
authors.initials: C
authors.lastname: Housset
authors.forename: P
authors.initials: P
authors.lastname: Balladur
authors.forename: H
authors.initials: H
authors.lastname: Toutain
authors.forename: V
authors.initials: V
authors.lastname: Albaladejo
mesh_terms: D000328:Adult; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005260:Female; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D008297:Male; D008875:Middle Aged; D009924:Organ Culture Techniques; D010100:Oxygen / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D003577:Cytochrome P-450 Enzyme System; D019392:Cytochrome P-450 CYP2E1; C104464:CYP3A protein, human; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D005978:Glutathione; D010100:Oxygen