Cell-Cell and Cell-Matrix Interactions Differentially Regulate the Expression of Hepatic and Cytoskeletal Genes in Primary Cultures of Rat Hepatocytes

Freshly isolated adult rat hepatocytes exhibit a flat, extended morphology when cultured on dried rat tail collagen in the presence of growth factors; they actively synthesize DNA and express high levels of cytoskeletal mRNAs and proteins (actin, tubulin, cytokeratins, vinculin, α -actinin, and desm...

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 1988-04, Vol.85 (7), p.2161-2165
Hauptverfasser: Avri Ben-Ze'ev, Robinson, Gregory S., Nancy L. R. Bucher, Farmer, Stephen R.
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container_title Proceedings of the National Academy of Sciences - PNAS
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creator Avri Ben-Ze'ev
Robinson, Gregory S.
Nancy L. R. Bucher
Farmer, Stephen R.
description Freshly isolated adult rat hepatocytes exhibit a flat, extended morphology when cultured on dried rat tail collagen in the presence of growth factors; they actively synthesize DNA and express high levels of cytoskeletal mRNAs and proteins (actin, tubulin, cytokeratins, vinculin, α -actinin, and desmoplakin), while exhibiting low levels of liver-specific mRNAs (albumin, α 1-inhibitor III, and α 1-antitrypsin) and limited synthesis and secretion of albumin. Hepatocytes cultured on hydrated gel matrix from the Engelbreth-Holm-Swarm (EHS) mouse tumor form small spherical aggregates and exhibit low DNA, cytoskeletal mRNA, and protein synthesis, while at the same time exhibiting elevated liver-specific mRNAs and albumin production; these cells, therefore, more nearly conform to the program of gene expression seen within the normal animal. Hepatocytes on hydrated rat tail collagen resemble those on dry collagen when cultured at low density, but at high density they form compact trabecular aggregates, synthesize negligible amounts of DNA, and maintain a pattern of gene expression resembling that of hepatocytes seeded on the EHS matrix. If cell morphology is compact, as on EHS or on hydrated rat tail collagen when densely populated, DNA synthesis and expression of cytoskeletal genes are low, while liver-specific mRNAs are abundant. When cells are extended the opposite is the case. Without the growth supplement DNA synthesis is low throughout but gene expression is little affected. These studies point to the importance of cell-cell and cell-matrix interactions in determining the differentiated phenotype of hepatocytes, and they reveal an inverse relationship between cytoskeletal and liver-specific protein expression.
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Hepatocytes cultured on hydrated gel matrix from the Engelbreth-Holm-Swarm (EHS) mouse tumor form small spherical aggregates and exhibit low DNA, cytoskeletal mRNA, and protein synthesis, while at the same time exhibiting elevated liver-specific mRNAs and albumin production; these cells, therefore, more nearly conform to the program of gene expression seen within the normal animal. Hepatocytes on hydrated rat tail collagen resemble those on dry collagen when cultured at low density, but at high density they form compact trabecular aggregates, synthesize negligible amounts of DNA, and maintain a pattern of gene expression resembling that of hepatocytes seeded on the EHS matrix. If cell morphology is compact, as on EHS or on hydrated rat tail collagen when densely populated, DNA synthesis and expression of cytoskeletal genes are low, while liver-specific mRNAs are abundant. When cells are extended the opposite is the case. Without the growth supplement DNA synthesis is low throughout but gene expression is little affected. 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R. Bucher</creatorcontrib><creatorcontrib>Farmer, Stephen R.</creatorcontrib><title>Cell-Cell and Cell-Matrix Interactions Differentially Regulate the Expression of Hepatic and Cytoskeletal Genes in Primary Cultures of Rat Hepatocytes</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Freshly isolated adult rat hepatocytes exhibit a flat, extended morphology when cultured on dried rat tail collagen in the presence of growth factors; they actively synthesize DNA and express high levels of cytoskeletal mRNAs and proteins (actin, tubulin, cytokeratins, vinculin, α -actinin, and desmoplakin), while exhibiting low levels of liver-specific mRNAs (albumin, α 1-inhibitor III, and α 1-antitrypsin) and limited synthesis and secretion of albumin. Hepatocytes cultured on hydrated gel matrix from the Engelbreth-Holm-Swarm (EHS) mouse tumor form small spherical aggregates and exhibit low DNA, cytoskeletal mRNA, and protein synthesis, while at the same time exhibiting elevated liver-specific mRNAs and albumin production; these cells, therefore, more nearly conform to the program of gene expression seen within the normal animal. Hepatocytes on hydrated rat tail collagen resemble those on dry collagen when cultured at low density, but at high density they form compact trabecular aggregates, synthesize negligible amounts of DNA, and maintain a pattern of gene expression resembling that of hepatocytes seeded on the EHS matrix. If cell morphology is compact, as on EHS or on hydrated rat tail collagen when densely populated, DNA synthesis and expression of cytoskeletal genes are low, while liver-specific mRNAs are abundant. When cells are extended the opposite is the case. Without the growth supplement DNA synthesis is low throughout but gene expression is little affected. These studies point to the importance of cell-cell and cell-matrix interactions in determining the differentiated phenotype of hepatocytes, and they reveal an inverse relationship between cytoskeletal and liver-specific protein expression.</description><subject>Albumins</subject><subject>Albumins - biosynthesis</subject><subject>Albumins - genetics</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell Communication</subject><subject>Cell culture techniques</subject><subject>Cell Differentiation</subject><subject>Collagen</subject><subject>Collagens</subject><subject>Contact Inhibition</subject><subject>Culture Media</subject><subject>Cultured cells</subject><subject>Cytoskeletal Proteins - biosynthesis</subject><subject>Cytoskeletal Proteins - genetics</subject><subject>DNA</subject><subject>DNA Replication</subject><subject>Extracellular Matrix - physiology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Hepatocytes</subject><subject>Liver - cytology</subject><subject>Liver - metabolism</subject><subject>Messenger RNA</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Organ Specificity</subject><subject>Rats</subject><subject>RNA, Messenger - biosynthesis</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptUctu1DAUtRCoDIUtCyQkLxC7hOvYie0lmpa2UhGogrXlca4hxZMMtiPN_AjfS0KGoQs217LO4z4OIS8ZlAwkf7frbSpVXcqyYg17RFYMNCsaoeExWQFUslCiEk_Js5TuAUDXCs7IGec151KsyK81hlDMhdq-pX9-H22O3Z7e9Bmjdbkb-kQvOu8xYp87G8KB3uG3MdiMNH9HernfRUxp4tHB02vc2dy5xe6Qh_QDA2Yb6BX2mGjX08-x29p4oOsx5HFSzqo7mxfl4A4Z03PyxNuQ8MXxPSdfP1x-WV8Xt5-ubtbvbwsntMxTlaC4aMRGMd7yGmTrtfCVY5VuGgGsVV4jWCVAyGYD2DRe165umXLCIefn5O3iu4vDzxFTNtsuuekItsdhTEYqVjOpm4lYLkQXh5QierNbtjAMzByEmYMwqjbSzEFMgtdH53GzxfZEP15-wt8ccZucDT7a3nXpRJOglWbswYCz_V_01Mb4MYSM-_yg33-JE_5qwe9THuK_cRgozX8DU8eywA</recordid><startdate>19880401</startdate><enddate>19880401</enddate><creator>Avri Ben-Ze'ev</creator><creator>Robinson, Gregory S.</creator><creator>Nancy L. 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Psychology</topic><topic>Gels</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>Hepatocytes</topic><topic>Liver - cytology</topic><topic>Liver - metabolism</topic><topic>Messenger RNA</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Organ Specificity</topic><topic>Rats</topic><topic>RNA, Messenger - biosynthesis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Avri Ben-Ze'ev</creatorcontrib><creatorcontrib>Robinson, Gregory S.</creatorcontrib><creatorcontrib>Nancy L. R. 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subjects Albumins
Albumins - biosynthesis
Albumins - genetics
Animals
Biological and medical sciences
Cell Communication
Cell culture techniques
Cell Differentiation
Collagen
Collagens
Contact Inhibition
Culture Media
Cultured cells
Cytoskeletal Proteins - biosynthesis
Cytoskeletal Proteins - genetics
DNA
DNA Replication
Extracellular Matrix - physiology
Fundamental and applied biological sciences. Psychology
Gels
Gene expression
Gene Expression Regulation
Hepatocytes
Liver - cytology
Liver - metabolism
Messenger RNA
Molecular and cellular biology
Molecular genetics
Organ Specificity
Rats
RNA, Messenger - biosynthesis
title Cell-Cell and Cell-Matrix Interactions Differentially Regulate the Expression of Hepatic and Cytoskeletal Genes in Primary Cultures of Rat Hepatocytes
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