Document Detail

3-D coculture of hepatic sinusoidal cells with primary hepatocytes-design of an organotypical model.
MedLine Citation:
PMID:  8660959     Owner:  NLM     Status:  MEDLINE    
Models for cocultures of parenchymal (PC) and nonparenchymal cells (NPC) of the liver relied on mixing the cells in a two-dimensional configuration or on establishing spheroidal aggregates. In vivo hepatic nonparenchymal cells, such as endothelial cells and Kupffer cells, are separated from parenchymal cells by extracellular matrix (ECM). Due to their location outside of the space of Disse they can form a barrier toward the sinusoid. Hepatocytes are attached to ECM of the space of Disse via two opposing sinusoidal surfaces. No three-dimensional coculture model reflecting this specific microenvironment of the liver cell plates in vivo has been available to date. We designed a three-dimensional model by positioning NPC on top of PC enclosed as a monolayer within a collagen sandwich. A gas-permeable membrane support can be used to allow the supply of oxygen to the resulting cell plate also from underneath the cell layers. Morphological analysis was performed by inverse and cross-sectional studies by light microscopy, scanning, and transmission electron microscopy of the coculture model. Cuboidal hepatocytes formed confluent layers below the NPC layer. They regularly expressed bile canaliculi at intercellular contact zones. Both sinusoidal surfaces expressed microprojections. Characteristic NPC including endothelial cells, Kupffer cells, and Ito cells completely covered the second matrix layer within a week. Kupffer cells were located on top of endothelial cells. Ito cells were intermingled and could be identified by their intracytoplasmic lipid droplets. LPS stimulation of cocultures resulted in a depression of albumin secretion. Phase I and phase II metabolites of the cytochrome P-450 1A1 substrate ethoxyresorufin were generated independently from the presence of cocultured NPC. This study describes the development of a novel three-dimensional coculture model, which intends to mimic more closely the microenvironment of the hepatic sinusoid by respecting the specific plate structure of the liver parenchyma. The model could serve as a complex tool to study potential collaborations between PC and NPC of the liver.
A Bader; E Knop; A Kern; K Böker; N Frühauf; O Crome; H Esselmann; C Pape; G Kempka; K F Sewing
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Experimental cell research     Volume:  226     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  1996 Jul 
Date Detail:
Created Date:  1996-08-22     Completed Date:  1996-08-22     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  223-33     Citation Subset:  IM    
Institut für Allgemeine Pharmakologie und Toxikologie, Medizinische Hochschule Hannover, Wuppertal, Germany.
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MeSH Terms
Albumins / secretion
Cell Culture Techniques / methods*
Cells, Cultured / cytology,  drug effects,  ultrastructure
Endothelium / cytology,  metabolism
Extracellular Matrix / metabolism
Lipopolysaccharides / pharmacology
Liver / cytology*
Microscopy, Electron
Oxazines / pharmacokinetics
Rats, Wistar
Reg. No./Substance:
0/Albumins; 0/Lipopolysaccharides; 0/Oxazines; 5725-91-7/ethoxyresorufin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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