Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1980 Nov;77(11):6616–6620. doi: 10.1073/pnas.77.11.6616

Control of serum protein production in hepatocyte hybridomas: immortalization and expression of normal hepatocyte genes.

J Szpirer, C Szpirer, J C Wanson
PMCID: PMC350337  PMID: 6161368

Abstract

"Hepatocyte hybridomas" have been isolated after fusion of adult hepatocytes and alpha-fetoprotein (AFP)-producing mouse hepatoma cells. The yield of viable hybrid clones was low but could be increased by culturing the cells in the presence of insulin. On the basis of their chromosome constitution, the hybrids were classified into two groups characterized by either a single or a double set of mouse (hepatoma) chromosomes. The hybrids segregated rat chromosomes and thus constitute an excellent material for gene mapping studies in the rat. Most of the hepatocyte hybridomas retained the production of one or more rat serum proteins, indicating that the corresponding structural genes, contributed by the normal hepatocyte parent, have been immortalized and maintained in the active state after fusion. However, these hybrids do not produce rat AFP, although mouse AFP synthesis is maintained. This result strongly suggests that silent rat (hepatocyte) AFP genes coexist in hepatocyte hybridoma nuclei with active mouse (hepatoma) AFP genes. Finally, on the basis of certain properties of these hepatocyte-hepatoma hybrids, we suggest that the nondividing state of the hepatocytes is actively controlled by a regulatory mechanism which prevents DNA synthesis or entry into mitosis or both.

Full text

PDF
6616

Images in this article

6618Image
on p.6618
6618Image
on p.6618
6618Image
on p.6618
6618Image
on p.6618
6618Image
on p.6618
6618Image
on p.6618
6618Image
on p.6618
6618Image
on p.6618

Selected References

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

  1. Abelev G. I. Alpha-fetoprotein in ontogenesis and its association with malignant tumors. Adv Cancer Res. 1971;14:295–358. doi: 10.1016/s0065-230x(08)60523-0. [DOI] [PubMed] [Google Scholar]
  2. Benedict W. F., Nebert D. W., Thompson E. B. Expression of aryl hydrocarbon hydroxylase induction and suppression of tyrosine aminotransferase induction in somatic-cell hybrids. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2179–2183. doi: 10.1073/pnas.69.8.2179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Crane L. J., Miller D. L. Plasma protein synthesis by isolated rat hepatocytes. J Cell Biol. 1977 Jan;72(1):11–25. doi: 10.1083/jcb.72.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Da Silva F. P., Hoecker G. F., Day N. K., Vienne K., Rubinstein P. Murine complement component 3: genetic variation and linkage to H-2. Proc Natl Acad Sci U S A. 1978 Feb;75(2):963–965. doi: 10.1073/pnas.75.2.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Deisseroth A., Nienhuis A., Turner P., Velez R., Anderson W. F., Ruddle F., Lawrence J., Creagan R., Kucherlapati R. Localization of the human alpha-globin structural gene to chromosome 16 in somatic cell hybrids by molecular hybridization assay. Cell. 1977 Sep;12(1):205–218. doi: 10.1016/0092-8674(77)90198-2. [DOI] [PubMed] [Google Scholar]
  6. Drochmans P., Wanson J. C., Mosselmans R. Isolation and subfractionation on ficoll gradients of adult rat hepatocytes. Size, morphology, and biochemical characteristics of cell fractions. J Cell Biol. 1975 Jul;66(1):1–22. doi: 10.1083/jcb.66.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gordon S., Cohn Z. Macrophage-melanocyte heterokaryons. I. Preparation and properties. J Exp Med. 1970 May 1;131(5):981–1003. doi: 10.1084/jem.131.5.981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gordon S., Ripps C. S., Cohn Z. The preparation and properties of macrophage-L cell hybrids. J Exp Med. 1971 Nov 1;134(5):1187–1200. doi: 10.1084/jem.134.5.1187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HAYFLICK L. THE LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS. Exp Cell Res. 1965 Mar;37:614–636. doi: 10.1016/0014-4827(65)90211-9. [DOI] [PubMed] [Google Scholar]
  10. Jeejeebhoy K. N., Ho J., Greenberg G. R., Phillips M. J., Bruce-Robertson A., Sodtke U. Albumin, fibrinogen and transferrin synthesis in isolated rat hepatocyte suspensions. A model for the study of plasma protein synthesis. Biochem J. 1975 Jan;146(1):141–155. doi: 10.1042/bj1460141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  12. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  13. Nichols E. A., Ruddle F. H., Petras M. L. Linkage of the locus for serum albumin in the house mouse, Mus musculus. Biochem Genet. 1975 Oct;13(9-10):551–555. doi: 10.1007/BF00484914. [DOI] [PubMed] [Google Scholar]
  14. Niwa A., Yamamoto K., Yasumura Y. Establishment of a rat hepatoma cell line which has ornithine carbamoyltransferase activity and grows continuously in arginine-deprived medium. J Cell Physiol. 1979 Jan;98(1):177–184. doi: 10.1002/jcp.1040980119. [DOI] [PubMed] [Google Scholar]
  15. Norwood T. H., Pendergrass W. R., Sprague C. A., Martin G. M. Dominance of the senescent phenotype in heterokaryons between replicative and post-replicative human fibroblast-like cells. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2231–2235. doi: 10.1073/pnas.71.6.2231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Riddle V. G., Harris H. Synthesis of a liver enzyme in hybrid cells. J Cell Sci. 1976 Oct;22(1):199–215. doi: 10.1242/jcs.22.1.199. [DOI] [PubMed] [Google Scholar]
  18. SHREFFLER D. C. LINKAGE OF THE MOUSE TRANSFERRIN LOCUS. J Hered. 1963 May-Jun;54:127–129. doi: 10.1093/oxfordjournals.jhered.a107241. [DOI] [PubMed] [Google Scholar]
  19. Schneider J. A., Weiss M. C. Expression of differentiated functions in hepatoma cell hybrids. I. Tyrosine aminotransferase in hepatoma-fibroblast hybrids. Proc Natl Acad Sci U S A. 1971 Jan;68(1):127–131. doi: 10.1073/pnas.68.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sell S., Becker F. F. alpha-Fetoprotein. J Natl Cancer Inst. 1978 Jan;60(1):19–26. doi: 10.1093/jnci/60.1.19. [DOI] [PubMed] [Google Scholar]
  21. Stein G. H., Yanishevsky R. M. Entry into S phase is inhibited in two immortal cell lines fused to senescent human diploid cells. Exp Cell Res. 1979 Apr;120(1):155–165. doi: 10.1016/0014-4827(79)90546-9. [DOI] [PubMed] [Google Scholar]
  22. Szpirer C., Szpirer J. A mouse hepatoma cell line which secretes several serum proteins including albumin and alpha-foetoprotein. Differentiation. 1975 Oct 16;4(2):85–91. doi: 10.1111/j.1432-0436.1975.tb01446.x. [DOI] [PubMed] [Google Scholar]
  23. Szpirer J., Szpirer C. Production of serum proteins in normal diploid fibroblast-hepatoma cell hybrids and in A9-normal liver cell hybrids. J Cell Sci. 1979 Feb;35:267–279. doi: 10.1242/jcs.35.1.267. [DOI] [PubMed] [Google Scholar]
  24. Szpirer J., Szpirer C. The control of serum protein synthesis in hepatoma-fibroblast hybrids. Cell. 1975 Sep;6(1):53–60. doi: 10.1016/0092-8674(75)90073-2. [DOI] [PubMed] [Google Scholar]
  25. Widman L. E., Golden J. J., Chasin L. A. Immortalization of normal liver functions in cell culture: rat hepatocyte-hepatoma cell hybrids expressing ornithine carbamoyltransferase activity. J Cell Physiol. 1979 Sep;100(3):391–400. doi: 10.1002/jcp.1041000302. [DOI] [PubMed] [Google Scholar]
  26. Yanishevsky R. M., Stein G. H. Ongoing DNA synthesis continues in young human diploid cells (HDC) fused to senescent HDC, but entry into S phase is inhibited. Exp Cell Res. 1980 Apr;126(2):469–472. doi: 10.1016/0014-4827(80)90290-6. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES