Modulation of protein synthesis and secretion by substratum in primary cultures of rat hepatocytes.
Hepatocytes isolated by perfusion of adult rat liver and cultured on substrata consisting of one or more of the major components of the liver biomatrix (fibronectin, laminin, type IV collagen) have been examined for the synthesis of defined proteins. Under these conditions, tyrosine amino transferase, a marker of hepatocyte function, is maintained at similar levels in response to dexamethasone over 5 days in culture on each substratum, and total cellular protein synthesis remains constant. By contrast, there is a rapid decrease in synthesis and secretion of albumin and a 3-7-fold increase in synthesis and secretion of alpha-fetoprotein which are most marked on a laminin substratum, but least evident on type IV collagen, and an increased synthesis of fibronectin and type IV collagen. The newly synthesized matrix proteins are present in the cell layer as well as in cell secretions. The enhanced synthesis of fibronectin is less in cells seeded onto a fibronectin substratum than on laminin or type IV collagen substrata, and its synthesis by hepatocytes seeded onto a mixed substratum of laminin and fibronectin is down-regulated by fibronectin in a dose-related manner. Similarly, type IV collagen synthesis is less when the cells are seeded on the homologous matrix protein substratum than on heterologous substrata. These results indicate that hepatocytes cultured in serum-free medium on substrata composed of components of the liver biomatrix maintain certain functions of the differentiated state (tyrosine amino transferase), lose others (albumin secretion) and switch to increased synthesis of matrix components as well as fetal markers such as alpha-fetoprotein. The magnitude of these effects depends on the substratum on which the hepatocytes are cultured.
D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D003470:Culture Media; D005353:Fibronectins / Q000096:biosynthesis; D008099:Liver / Q000378:metabolism*; D014176:Protein Biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D012709:Serum Albumin / Q000096:biosynthesis; D000509:alpha-Fetoproteins / Q000096:biosynthesis
D003470:Culture Media; D005353:Fibronectins; D012709:Serum Albumin; D000509:alpha-Fetoproteins; D003094:Collagen