Reversal of activation of human myofibroblast-like cells by culture on a basement membrane-like substrate.
BACKGROUND
Liver injury transforms hepatic stellate cells into myofibroblast (MFB)-like cells. With recovery from injury, MFBs undergo apoptosis, but it is unknown whether they can also revert to quiescence.


OBJECTIVE
To determine whether human (h)MFBs become quiescent if cultured on a basement membrane-like substrate (Matrigel).


METHODS
hMFBs obtained from cirrhotic liver were re-cultured on plastic or Matrigel. Expression of genes of collagen metabolism was assayed before and after transforming growth factor beta (TGFbeta) and Oncostatin M (OSM) stimulation.


RESULTS
hMFBs had typical MFB-like morphology, with abundant alpha-smooth muscle actin (SMA) but no cytoplasmic lipid droplets. hMFBs re-cultured on Matrigel reverted to alphaSMA-negative, lipid droplet-positive quiescent morphology. alphaSMA, collagen alpha1(1) (COL1A1) and collagen alpha2(1) (COL1A2) messages were upregulated in hMFBs cultured on plastic, but suppressed by Matrigel. The opposite was true for metalloproteinase-1 mRNA. OSM but not TGFbeta reduced alphaSMA mRNA by 30% while TGFbeta but not OSM upregulated COL1A1 mRNA by 48%, in hMFBs on plastic. TGFbeta and OSM stimulated COL1A1 gene expression in Matrigel by 50 and 60%, respectively.


CONCLUSIONS
Matrigel culture de-activates hMFBs yet collagen gene expression still responds to fibrogenic cytokines. The responses of hMFB gene expression to TGFbeta and OSM, are regulated differently by the extracellular matrix.
Division of Gastroenterology and Hepatology, Medical University of South Carolina, 96 Jonathan Lucas St., Suite 210, Charleston 29425, USA.
Naondo
N
Sohara
Iya
I
Znoyko
Miriam T
MT
Levy
Maria
M
Trojanowska
Adrian
A
Reuben
D000199:Actins / Q000235:genetics; D000970:Antineoplastic Agents / Q000494:pharmacology; D001485:Basement Membrane; D001672:Biocompatible Materials / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics / Q000494:pharmacology; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D004338:Drug Combinations; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology*; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D020781:Matrix Metalloproteinase 1 / Q000235:genetics; D053683:Oncostatin M; D010455:Peptides / Q000494:pharmacology; D010969:Plastics; D011509:Proteoglycans / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D000199:Actins; D000970:Antineoplastic Agents; D001672:Biocompatible Materials; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D004338:Drug Combinations; D007797:Laminin; C508351:OSM protein, human; D010455:Peptides; D010969:Plastics; D011509:Proteoglycans; D012333:RNA, Messenger; D019715:Tissue Inhibitor of Metalloproteinase-1; D016212:Transforming Growth Factor beta; D053683:Oncostatin M; C059485:matrigel; D003094:Collagen; D020781:Matrix Metalloproteinase 1