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. 1983 Dec;80(23):7229–7233. doi: 10.1073/pnas.80.23.7229

Reciprocal modulation of growth and differentiated functions of mature rat hepatocytes in primary culture by cell--cell contact and cell membranes.

T Nakamura, K Yoshimoto, Y Nakayama, Y Tomita, A Ichihara
PMCID: PMC390028  PMID: 6580642

Abstract

In primary monolayer cultures of rat mature hepatocytes, many metabolic functions as well as cell growth are regulated by cell density. There are two types of regulatory response of these functions to change of cell density. Growth-related functions, such as DNA synthesis, induction of glucose-6-phosphate dehydrogenase, 2-aminoisobutyric acid transport, synthesis of cellular protein, and cholesterogenesis, are stimulated by low cell density. In contrast, functions related to hepatocyte-specific characters, such as the inductions of tyrosine aminotransferase, serine dehydratase, and malic enzyme and synthesis of triglycerides, are stimulated by high cell density. The reciprocal responses of these cellular activities to cell density were mimicked by addition of plasma membranes purified from adult rat liver to hepatocytes cultured at low cell density. The modulator activity was heat labile and trypsin sensitive. The activity was also found in plasma membranes from kidney, brain, and erythrocytes, although the specific activities of these preparations seemed to be different. These results suggest that the reciprocal regulations of cell growth and hepatocyte-specific functions are mediated by some surface components via cell-cell contact.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Chung S., Landfear S. M., Blumberg D. D., Cohen N. S., Lodish H. F. Synthesis and stability of developmentally regulated dictyostelium mRNAs are affected by cell--cell contact and cAMP. Cell. 1981 Jun;24(3):785–797. doi: 10.1016/0092-8674(81)90104-5. [DOI] [PubMed] [Google Scholar]
  4. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  5. Hausman R. E., Moscona A. A. Isolation of retina-specific cell-aggregating factor from membranes of embryonic neural retina tissue. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3594–3598. doi: 10.1073/pnas.73.10.3594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Holley R. W., Böhlen P., Fava R., Baldwin J. H., Kleeman G., Armour R. Purification of kidney epithelial cell growth inhibitors. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5989–5992. doi: 10.1073/pnas.77.10.5989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Holley R. W., Kiernan J. A. "Contact inhibition" of cell division in 3T3 cells. Proc Natl Acad Sci U S A. 1968 May;60(1):300–304. doi: 10.1073/pnas.60.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ichihara A., Nakamura T., Tanaka K. Use of hepatocytes in primary culture for biochemical studies on liver functions. Mol Cell Biochem. 1982 Apr 2;43(3):145–160. doi: 10.1007/BF00223006. [DOI] [PubMed] [Google Scholar]
  9. Jones D. H., Matus A. I. Isolation of synaptic plasma membrane from brain by combined flotation-sedimentation density gradient centrifugation. Biochim Biophys Acta. 1974 Aug 9;356(3):276–287. doi: 10.1016/0005-2736(74)90268-5. [DOI] [PubMed] [Google Scholar]
  10. Kaleko M., Rothman F. G. Membrane sites regulating developmental gene expression in Dictyostelium discoideum. Cell. 1982 Apr;28(4):801–811. doi: 10.1016/0092-8674(82)90059-9. [DOI] [PubMed] [Google Scholar]
  11. Kletzien R. F., Pariza M. W., Becker J. E., Potter V. R., Butcher F. R. Induction of amino acid transport in primary cultures of adult rat liver parenchymal cells by insulin. J Biol Chem. 1976 May 25;251(10):3014–3020. [PubMed] [Google Scholar]
  12. McGowan J. A., Strain A. J., Bucher N. L. DNA synthesis in primary cultures of adult rat hepatocytes in a defined medium: effects of epidermal growth factor, insulin, glucagon, and cyclic-AMP. J Cell Physiol. 1981 Sep;108(3):353–363. doi: 10.1002/jcp.1041080309. [DOI] [PubMed] [Google Scholar]
  13. Meyer D. J., Yancey S. B., Revel J. P. Intercellular communication in normal and regenerating rat liver: a quantitative analysis. J Cell Biol. 1981 Nov;91(2 Pt 1):505–523. doi: 10.1083/jcb.91.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Michalopoulos G., Cianciulli H. D., Novotny A. R., Kligerman A. D., Strom S. C., Jirtle R. L. Liver regeneration studies with rat hepatocytes in primary culture. Cancer Res. 1982 Nov;42(11):4673–4682. [PubMed] [Google Scholar]
  15. Morris J. E., Moscona A. A. The induction of glutamine synthetase in cell aggregates of embryonic neural retina: correlations with differentation and multicellular organization. Dev Biol. 1971 Jul;25(3):420–444. doi: 10.1016/0012-1606(71)90040-6. [DOI] [PubMed] [Google Scholar]
  16. Nakamura T., Aoyama K., Tomomura A., Ichihara A. Hormonal control of the development of tryptophan oxygenase in primary cultures of young rat hepatocytes. Biochim Biophys Acta. 1981 Nov 18;678(1):91–97. [PubMed] [Google Scholar]
  17. Nakamura T., Shinno H., Ichihara A. Insulin and glucagon as a new regulator system for tryptophan oxygenase activity demonstrated in primary cultured rat hepatocytes. J Biol Chem. 1980 Aug 25;255(16):7533–7535. [PubMed] [Google Scholar]
  18. Nakamura T., Tomita Y., Ichihara A. Density-dependent growth control of adult rat hepatocytes in primary culture. J Biochem. 1983 Oct;94(4):1029–1035. doi: 10.1093/oxfordjournals.jbchem.a134444. [DOI] [PubMed] [Google Scholar]
  19. Nakamura T., Tomomura A., Noda C., Shimoji M., Ichihara A. Acquisition of a beta-adrenergic response by adult rat hepatocytes during primary culture. J Biol Chem. 1983 Aug 10;258(15):9283–9289. [PubMed] [Google Scholar]
  20. Nakamura T., Yoshimoto K., Aoyama K., Ichihara A. Hormonal regulations of glucose-6-phosphate dehydrogenase and lipogenesis in primary cultures of rat hepatocytes. J Biochem. 1982 Feb;91(2):681–693. doi: 10.1093/oxfordjournals.jbchem.a133741. [DOI] [PubMed] [Google Scholar]
  21. Noda C., Nakamura T., Ichihara A. alpha-Adrenergic regulation of enzymes of amino acid metabolism in primary cultures of adult rat hepatocytes. J Biol Chem. 1983 Feb 10;258(3):1520–1525. [PubMed] [Google Scholar]
  22. Okamoto K., Takeuchi I. Changes in activities of two developmentally regulated enzymes induced by disaggregation of the pseudoplasmodia of Dictyostelium discoideum. Biochem Biophys Res Commun. 1976 Sep 20;72(2):739–746. doi: 10.1016/s0006-291x(76)80101-5. [DOI] [PubMed] [Google Scholar]
  23. Richman R. A., Claus T. H., Pilkis S. J., Friedman D. L. Hormonal stimulation of DNA synthesis in primary cultures of adult rat hepatocytes. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3589–3593. doi: 10.1073/pnas.73.10.3589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sacktor B., Rosenbloom I. L., Liang C. T., Cheng L. Sodium gradient- and sodium plus potassium gradient-dependent L-glutamate uptake in renal basolateral membrane vesicles. J Membr Biol. 1981 May 15;60(1):63–71. doi: 10.1007/BF01870833. [DOI] [PubMed] [Google Scholar]
  25. Seglen P. O. Preparation of isolated rat liver cells. Methods Cell Biol. 1976;13:29–83. doi: 10.1016/s0091-679x(08)61797-5. [DOI] [PubMed] [Google Scholar]
  26. Steck P. A., Blenis J., Voss P. G., Wang J. L. Growth control in cultured 3T3 fibroblasts II. Molecular properties of a fraction enriched in growth inhibitory activity. J Cell Biol. 1982 Feb;92(2):523–530. doi: 10.1083/jcb.92.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stoker M. G. Role of diffusion boundary layer in contact inhibition of growth. Nature. 1973 Nov 23;246(5430):200–203. doi: 10.1038/246200a0. [DOI] [PubMed] [Google Scholar]
  28. Tanaka K., Sato M., Tomita Y., Ichihara A. Biochemical studies on liver functions in primary cultured hepatocytes of adult rats. I. Hormonal effects on cell viability and protein synthesis. J Biochem. 1978 Oct;84(4):937–946. doi: 10.1093/oxfordjournals.jbchem.a132207. [DOI] [PubMed] [Google Scholar]
  29. Thiery J. P., Brackenbury R., Rutishauser U., Edelman G. M. Adhesion among neural cells of the chick embryo. II. Purification and characterization of a cell adhesion molecule from neural retina. J Biol Chem. 1977 Oct 10;252(19):6841–6845. [PubMed] [Google Scholar]
  30. Tomita Y., Nakamura T., Ichihara A. Control of DNA synthesis and ornithine decarboxylase activity by hormones and amino acids in primary cultures of adult rat hepatocytes. Exp Cell Res. 1981 Oct;135(2):363–371. doi: 10.1016/0014-4827(81)90172-5. [DOI] [PubMed] [Google Scholar]
  31. WISE E. M., Jr, BALL E. G. MALIC ENZYME AND LIPOGENESIS. Proc Natl Acad Sci U S A. 1964 Nov;52:1255–1263. doi: 10.1073/pnas.52.5.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Whittenberger B., Glaser L. Inhibition of DNA synthesis in cultures of 3T3 cells by isolated surface membranes. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2251–2255. doi: 10.1073/pnas.74.6.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Whittenberger B., Raben D., Lieberman M. A., Glaser L. Inhibition of growth of 3T3 cells by extract of surface membranes. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5457–5461. doi: 10.1073/pnas.75.11.5457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yee A. G., Revel J. P. Loss and reappearance of gap junctions in regenerating liver. J Cell Biol. 1978 Aug;78(2):554–564. doi: 10.1083/jcb.78.2.554. [DOI] [PMC free article] [PubMed] [Google Scholar]

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