PMID- 17361319
OWN - NLM
STAT- MEDLINE
DCOM- 20070626
LR  - 20191210
IS  - 1868-596X (Print)
IS  - 1868-596X (Linking)
VI  - 24
IP  - 1
DP  - 2007
TI  - Use of a standardised and validated long-term human hepatocyte culture system for 
      repetitive analyses of drugs: repeated administrations of acetaminophen reduces 
      albumin and urea secretion.
PG  - 35-40
AB  - Human hepatocytes are the in vitro system of choice to study drug-induced 
      processes in man. Here, we present HEPAC(2): a standardised and validated culture 
      system in which human hepatocytes are maintained in HHMM (Human Hepatocyte 
      Maintenance Medium) with HGF (hepatocyte growth factor) and EGF (epidermal growth 
      factor). Cellular viability and hepatocellular functions were monitored daily. 
      Albumin and urea production remained on a relatively constant level for up to 2-3 
      weeks. Based on this, a standard protocol was established that allows repeated 
      exposure of hepatocytes to study drug metabolism. We used acetaminophen (AAP) to 
      assay the feasibility of this system. Hepatocytes were exposed to AAP (100-2815 
      mg/l) for 24 h. Subsequently, the culture medium was replaced by medium without 
      AAP and the same exposure scenario was repeated at intervals of 4 days. High 
      doses of AAP (2815 mg/l) diminished urea production by 15-30% and albumin 
      secretion by 70-80%. These effects were reversible. After removal of AAP, 
      secretion of urea and albumin returned to control levels. AAP hepatotoxicity is 
      caused by its biotransformation to the reactive metabolite 
      N-acetyl-p-benzoquinoneimine (NAPQI) mediated by CYP2E1 and CYP1A2. The AAP 
      activating enzymes were active for at least 21 days and their activity was 
      maintained during at least four repeated cycles of exposure to AAP. In 
      conclusion, these data demonstrate the suitability of our long-term culture 
      system to serve as a tool for repetitive screening of drug-mediated changes of 
      hepatocellular functions. This culture technique may help to overcome the sparse 
      availability of human hepatocytes for testing drugmediated responses in man.
FAU - Ullrich, Anett
AU  - Ullrich A
AD  - PRIMACYT Cell Culture Technology GmbH, FRG, Schwerin, Germany.
FAU - Berg, Christine
AU  - Berg C
FAU - Hengstler, Jan G
AU  - Hengstler JG
FAU - Runge, Dieter
AU  - Runge D
LA  - eng
PT  - Journal Article
PT  - Validation Study
PL  - Germany
TA  - ALTEX
JT  - ALTEX
JID - 100953980
RN  - 0 (Albumins)
RN  - 0 (Analgesics, Non-Narcotic)
RN  - 362O9ITL9D (Acetaminophen)
RN  - 8W8T17847W (Urea)
RN  - EC 1.14.13.- (Cytochrome P-450 CYP2E1)
RN  - EC 1.14.14.1 (Cytochrome P-450 CYP1A2)
SB  - IM
MH  - Acetaminophen/*toxicity
MH  - Albumins/*drug effects/metabolism
MH  - Analgesics, Non-Narcotic/*toxicity
MH  - *Animal Testing Alternatives
MH  - Animals
MH  - Cells, Cultured
MH  - Cytochrome P-450 CYP1A2/drug effects/metabolism
MH  - Cytochrome P-450 CYP2E1/drug effects/metabolism
MH  - Dose-Response Relationship, Drug
MH  - Humans
MH  - Liver/cytology/*drug effects
MH  - Urea/*metabolism
EDAT- 2007/03/16 09:00
MHDA- 2007/06/27 09:00
CRDT- 2007/03/16 09:00
PHST- 2007/03/16 09:00 [pubmed]
PHST- 2007/06/27 09:00 [medline]
PHST- 2007/03/16 09:00 [entrez]
AID - 10.14573/altex.2007.1.35 [doi]
PST - ppublish
SO  - ALTEX. 2007;24(1):35-40. doi: 10.14573/altex.2007.1.35.