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Characterization of spontaneous cell death in monolayer cultures of primary hepatocytes

Mathieu Vinken, Elke Decrock UGent, Tatyana Doktorova, Eva Ramboer, Elke De Vuyst UGent, Tamara Vanhaecke, Luc Leybaert UGent and Vera Rogiers (2011) ARCHIVES OF TOXICOLOGY. 85(12). p.1589-1596
abstract
Monolayer cultures of primary hepatocytes, isolated from freshly removed livers, represent widely used in vitro tools in the area of liver physiology and pathology, pharmacology and toxicology. However, a major shortcoming of these systems is that they cope with dedifferentiation, which is accompanied by spontaneous cell death. The goal of the present study was to elucidate the mechanisms that drive the process of self-generated cell demise in primary hepatocyte cultures. For this purpose, isolated rat hepatocytes were cultivated under conventional conditions, and the occurrence of apoptosis and necrosis was monitored during 4 days by performing a set of acknowledged cell death assays. These included examination of cell morphology by light microscopy, quantification of apoptotic and necrotic cell populations by Hoechst 33342 and propidium iodide in situ staining, assessment of apoptotic and necrotic activities by measuring caspase 3-like activity and extracellular leakage of lactate dehydrogenase, and studying the expression of apoptosis regulators through immunoblot analysis. In essence, two cell death peaks were observed, namely shortly after cell seeding and in the final stages of the cultivation period, both involving apoptotic and necrotic actions. The outcome of this study not only sheds new light onto the molecular processes that underlie spontaneous cell death in primary hepatocyte cultures, but also opens perspectives for the establishment of strategies to increase cell survival in these popular in vitro systems.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Spontaneous apoptosis, Primary hepatocytes, Secondary necrosis, RAT HEPATOCYTES, SPONTANEOUS APOPTOSIS, MOLECULAR-MECHANISMS, SIGNALING PATHWAYS, SANDWICH CULTURES, TRICHOSTATIN-A, LIVER-INJURY, IN-VITRO, DEXAMETHASONE, NECROSIS
journal title
ARCHIVES OF TOXICOLOGY
Arch. Toxicol.
volume
85
issue
12
pages
1589 - 1596
Web of Science type
Article
Web of Science id
000297359700013
JCR category
TOXICOLOGY
JCR impact factor
4.674 (2011)
JCR rank
8/83 (2011)
JCR quartile
1 (2011)
ISSN
0340-5761
DOI
10.1007/s00204-011-0703-4
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2106397
handle
http://hdl.handle.net/1854/LU-2106397
date created
2012-05-14 20:50:01
date last changed
2012-05-15 10:03:38
@article{2106397,
  abstract     = {Monolayer cultures of primary hepatocytes, isolated from freshly removed livers, represent widely used in vitro tools in the area of liver physiology and pathology, pharmacology and toxicology. However, a major shortcoming of these systems is that they cope with dedifferentiation, which is accompanied by spontaneous cell death. The goal of the present study was to elucidate the mechanisms that drive the process of self-generated cell demise in primary hepatocyte cultures. For this purpose, isolated rat hepatocytes were cultivated under conventional conditions, and the occurrence of apoptosis and necrosis was monitored during 4 days by performing a set of acknowledged cell death assays. These included examination of cell morphology by light microscopy, quantification of apoptotic and necrotic cell populations by Hoechst 33342 and propidium iodide in situ staining, assessment of apoptotic and necrotic activities by measuring caspase 3-like activity and extracellular leakage of lactate dehydrogenase, and studying the expression of apoptosis regulators through immunoblot analysis. In essence, two cell death peaks were observed, namely shortly after cell seeding and in the final stages of the cultivation period, both involving apoptotic and necrotic actions. The outcome of this study not only sheds new light onto the molecular processes that underlie spontaneous cell death in primary hepatocyte cultures, but also opens perspectives for the establishment of strategies to increase cell survival in these popular in vitro systems.},
  author       = {Vinken, Mathieu and Decrock, Elke and Doktorova, Tatyana and Ramboer, Eva and De Vuyst, Elke and Vanhaecke, Tamara and Leybaert, Luc and Rogiers, Vera},
  issn         = {0340-5761},
  journal      = {ARCHIVES OF TOXICOLOGY},
  keyword      = {Spontaneous apoptosis,Primary hepatocytes,Secondary necrosis,RAT HEPATOCYTES,SPONTANEOUS APOPTOSIS,MOLECULAR-MECHANISMS,SIGNALING PATHWAYS,SANDWICH CULTURES,TRICHOSTATIN-A,LIVER-INJURY,IN-VITRO,DEXAMETHASONE,NECROSIS},
  language     = {eng},
  number       = {12},
  pages        = {1589--1596},
  title        = {Characterization of spontaneous cell death in monolayer cultures of primary hepatocytes},
  url          = {http://dx.doi.org/10.1007/s00204-011-0703-4},
  volume       = {85},
  year         = {2011},
}

Chicago
Vinken, Mathieu, Elke Decrock, Tatyana Doktorova, Eva Ramboer, Elke De Vuyst, Tamara Vanhaecke, Luc Leybaert, and Vera Rogiers. 2011. “Characterization of Spontaneous Cell Death in Monolayer Cultures of Primary Hepatocytes.” Archives of Toxicology 85 (12): 1589–1596.
APA
Vinken, M., Decrock, E., Doktorova, T., Ramboer, E., De Vuyst, E., Vanhaecke, T., Leybaert, L., et al. (2011). Characterization of spontaneous cell death in monolayer cultures of primary hepatocytes. ARCHIVES OF TOXICOLOGY, 85(12), 1589–1596.
Vancouver
1.
Vinken M, Decrock E, Doktorova T, Ramboer E, De Vuyst E, Vanhaecke T, et al. Characterization of spontaneous cell death in monolayer cultures of primary hepatocytes. ARCHIVES OF TOXICOLOGY. 2011;85(12):1589–96.
MLA
Vinken, Mathieu, Elke Decrock, Tatyana Doktorova, et al. “Characterization of Spontaneous Cell Death in Monolayer Cultures of Primary Hepatocytes.” ARCHIVES OF TOXICOLOGY 85.12 (2011): 1589–1596. Print.