Hepatocyte survival depends on beta1-integrin-mediated attachment of hepatocytes to hepatic extracellular matrix.
BACKGROUND
A major drawback of allogeneic hepatocyte transplantation is the lack of sustained survival of the transplanted cells in the recipient liver parenchyma. The purpose of this study was to determine the effect of the presence or absence of hepatic extracellular matrix (ECM) molecules on hepatocyte survival and function following hepatocyte isolation for transplantation purposes, and the role of beta1-integrin molecules therein.


METHODS
Hepatocytes, either untreated or treated with anti-beta1 integrin antibodies or RGD peptides, were seeded on wells precoated with collagen type I, type IV, laminin, fibronectin or polyhydroxyethylmehacrylate. The extent of attachment and apoptosis was evaluated.


RESULTS
When hepatocytes were added into wells precoated with either fibronectin, or collagen type IV, rapid spreading and prolonged survival occurred, in contrast to hepatocytes that were seeded in wells precoated with collagen type I or polyhydroxyethylmehacrylate. Pretreatment of the cells with anti-beta1-integrin antibodies resulted in reduction of cell attachment to laminin, fibronectin, collagen I, and collagen IV. Synthetic RGD (arginine-glycine-aspartate)-peptides and anti-beta1 antibodies inhibited apoptosis of cultured hepatocytes.


CONCLUSIONS
Our findings indicate that embedding of hepatocytes within their normal liver ECM surroundings maintains their survival. When detached from their natural surrounding hepatocytes enter into apoptosis, unless treated with anti-beta1-integrin antibodies or RGD peptides. This knowledge will allow improvement of hepatocyte transplantation efficiency.
Pinkse
Gabriëlle G M
GG
Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands. G.Pinkse@lumc.nl
Voorhoeve
Mathijs P
MP
Noteborn
Mathieu
M
Terpstra
Onno T
OT
Bruijn
Jan Anthonie
JA
De Heer
Emile
E
D000818:Animals; D023102:Anoikis; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000494:pharmacology; D053938:DNA Fragmentation; D004586:Electrophoresis; D005109:Extracellular Matrix / Q000502:physiology*; D005260:Female; D005353:Fibronectins / Q000494:pharmacology; D022781:Hepatocytes / Q000502:physiology*; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000276:immunology / Q000502:physiology*; D008099:Liver / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D051381:Rats; D011914:Rats, Inbred BN
D000911:Antibodies, Monoclonal; D024141:Collagen Type IV; D005353:Fibronectins; D019012:Integrin beta1; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases