+ Translate

Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix

, : Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix. Journal of Cellular Physiology 134(3): 309-323

Freshly isolated adult rat hepatocytes, when cultured on type I collagen (commercially available as Vitrogen), assume a polygonal shape, form a stable monolayer within 24 hours, but lose the capacity to express some liver-specific functions over time in culture. We incubated hepatocytes in a serum-free medium on a reconstituted basement membrane gel, "matrigel" (prepared from an extract of extracellular matrix of the murine Engelbreth-Holm-Swarm sarcoma), and observed that the cells adhered firmly, remained rounded as single cells or clusters, and maintained liver-specific gene expression for more than 1 week in vitro. Hepatocytes on matrigel secreted substantially higher amounts of albumin, transferrin, haptoglobin, and hemopexin, contained higher amounts of glutathione peroxidase, and, as judged by Northern blot analyses of extracted cellular RNA, expressed increased amounts of mRNA for the liver-specific protein albumin (as compared with cells on vitrogen). In cultures treated with phenobarbital, cytochrome P-450b, and cytochrome P-450e, mRNAs and proteins were barely detectable in cells on Vitrogen but were induced to levels similar to those in the liver in vivo in matrigel cultures. Likewise, the use of matrigel greatly enhanced the induction of mRNA and protein for P-450c by 3-methylcholanthrene and for P-450p by steroidal and nonsteroidal inducers. However, neither substratum permitted induction of P-450d by 3-methylcholanthrene, suggesting that the effects of matrigel are selective even for expression in liver of members of the superfamily of cytochrome P-450 genes. Within 5 days in cultures on Vitrogen, hepatocytes expressed detectable amounts of fetal liver aldolase activity and also mRNA for vimentin and type I collagen, each considered a phenotypic change reflecting hepatocyte "dedifferentiation". None of these was present in cells on matrigel. Responsiveness to mitogenic stimuli, as judged by incorporation of 3H-thymidine into DNA, was also decreased in hepatocytes cultured on matrigel. Finally, there was a remarkable increase in the levels of mRNAs for the cytoskeleton proteins actin and tubulin in hepatocytes on both matrices during the first 2 days in culture. However, the continuously flattening hepatocytes on Vitrogen maintained substantially higher levels of cytoskeleton mRNA over time in culture than did the rounded cells on matrigel. We conclude that hepatocytes cultured on matrigel, as opposed to the standard collagen, exhibit remarkably enhanced expression of many liver-specific functions.

Accession: 006288002

PMID: 3350857

DOI: 10.1002/jcp.1041340302

Download PDF Full Text: Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix

Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:

Related references

Schuetz, E.G.; Schuetz, J.D.; May, B.; Guzelian, P.S., 1990: Regulation of cytochrome P-450b/e and P-450p gene expression by growth hormone in adult rat hepatocytes cultured on a reconstituted basement membrane. Growth hormone (GH) may directly affect the expression of some liver cytochromes P-450 inducible by xenobiotics, but this has been difficult to establish with pituitary ablation in living animals. Therefore, we incubated adult rat hepatocytes on a...

Bissell, D.M.; Caron, J.M.; Babiss, L.E.; Friedman, J.M., 1990: Transcriptional regulation of the albumin gene in cultured rat hepatocytes. Role of basement-membrane matrix. The expression of tissue-specific functions by hepatocytes in primary culture is enhanced in the presence of an extracellular matrix. A basement membrane-like substratum, derived from the Engelbreth-Holm-Swarm mouse sarcoma (EHS) and termed EHS ge...

Schuetz E.G.; Schuetz J.; L.D.; Mode A.; Gustafsson J.A.; Guzelian P.S., 1988: Growth hormone regulation of sex specific and phenobarbital inducible cytochromes p 450 in adult rat hepatocytes cultured on the basement membrane matrix matrigel. Hepatology 8(5): 1227

Guzelian P., 1986: Regulation of gene expression in adult rat hepatocytes cultured on extracellular matrix. Hepatology: 1188

Aggeler, J.; Park, C.S.; Bissell, M.J., 1988: Regulation of milk protein and basement membrane gene expression: the influence of the extracellular matrix. Synthesis and secretion of milk proteins (.alpha.-casein, .beta.-casein, .gamma.-casein, and transferrin) by cultured primary mouse mammary epithelial cells is modulated by the extracellular matrix. In cells grown on released or "floating&quo...

Vandenberghe, Y.; Tee, L.; Morel, F.; Rogiers, V.; Guillouzo, A.; Yeoh, G., 1991: Regulation of glutathione S-transferase gene expression by phenobarbital in cultured adult rat hepatocytes. Previous studies, by using Northern blotting analyses, showed that phenobarbital (PB) affects the steady-state mRNA levels of glutathione S-transferase (GST) subunits 1/2, 3/4 and 7 in both conventional cultures of adult rat hepatocytes and co-cul...

Brill, S.; Zvibel, I.; Halpern, Z.; Oren, R., 2002: The role of fetal and adult hepatocyte extracellular matrix in the regulation of tissue-specific gene expression in fetal and adult hepatocytes. We explored the effect of extracellular matrix (ECM) produced by fetal and adult hepatocytes on tissue-specific gene expression and proliferation of fetal and adult hepatocytes. Adult hepatocytes ECM strongly induced expression of both albumin and...

Luttringer, O.; Theil, F.P.ter; Lave, T.; Wernli Kuratli, K.; Guentert, T.W.; D.S.izieu, A., 2002: Influence of isolation procedure, extracellular matrix and dexamethasone on the regulation of membrane transporters gene expression in rat hepatocytes. The influence of the isolation procedure of hepatocytes, extracellular matrix (ECM) configuration and incubation medium supplementation by dexamethasone (DEX) on the cell morphology and on the gene expression of membrane transporters was examined...

Wang, H.; Gao, X.; Tanihata, T.; Fukumoto, S.; Hirai, K., 1997: Transient expression and regulation of inducible nitric oxide synthase gene in primary cultured adult murine hepatocytes. Japanese Journal of Pharmacology 75(SUPPL 1): 84P

Clement B.; Rescan P.Y.; Loreal O.; Yamada Y.; Hassell J.R.; Guillouzo A., 1990: Differential expression of basement membrane genes in fetal and adult hepatocytes. Journal of Cell Biology 111(5 PART 2): 399A