Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1993 May;142(5):1373–1382.

Regulation of the differentiation of diploid and some aneuploid rat liver epithelial (stemlike) cells by the hepatic microenvironment.

W B Coleman 1, A E Wennerberg 1, G J Smith 1, J W Grisham 1
PMCID: PMC1886923  PMID: 8494041

Abstract

Following intrahepatic transplantation in adult syngeneic Fischer 344 rats, diploid cultured rat liver epithelial cells (WB-F344), modified to carry the Escherichia coli beta-galactosidase reporter gene and/or the fluorescent membrane dye PKH26-GL, integrate into hepatic plates and acquire the size and nuclear structure of mature hepatocytes. Additionally, of two aneuploid, neoplastically transformed derivatives of WB-F344 cells, both of which produce aggressively growing tumors when transplanted subcutaneously, cells of one line (GN6TF) do not produce tumors in the liver but integrate into hepatic plates and morphologically differentiate. The other transformed line (GP7TB) retains tumorigenicity in the liver, but cells in the intrahepatic tumors are more differentiated morphologically than are tumors at subcutaneous sites. These results suggest that WB-F344 cells are stemlike cells for hepatocytes and that the hepatic microenvironment induces them to incorporate into hepatic plates and differentiate. Our results also suggest that the hepatic microenvironment regulates the differentiation of some neoplastically transformed hepatic stemlike cells, thereby eliminating or reducing their tumorigenic potential.

Full text

PDF
1373

Images in this article

1376Figure 1
on p.1376
1377Figure 2
on p.1377
1378Figure 3
on p.1378
1379Figure 4
on p.1379

Selected References

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

  1. Aterman K. The stem cells of the liver--a selective review. J Cancer Res Clin Oncol. 1992;118(2):87–115. doi: 10.1007/BF01187498. [DOI] [PubMed] [Google Scholar]
  2. Beck E., Ludwig G., Auerswald E. A., Reiss B., Schaller H. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene. 1982 Oct;19(3):327–336. doi: 10.1016/0378-1119(82)90023-3. [DOI] [PubMed] [Google Scholar]
  3. Cepko C. L., Roberts B. E., Mulligan R. C. Construction and applications of a highly transmissible murine retrovirus shuttle vector. Cell. 1984 Jul;37(3):1053–1062. doi: 10.1016/0092-8674(84)90440-9. [DOI] [PubMed] [Google Scholar]
  4. Evarts R. P., Nagy P., Marsden E., Thorgeirsson S. S. A precursor-product relationship exists between oval cells and hepatocytes in rat liver. Carcinogenesis. 1987 Nov;8(11):1737–1740. doi: 10.1093/carcin/8.11.1737. [DOI] [PubMed] [Google Scholar]
  5. Evarts R. P., Nagy P., Nakatsukasa H., Marsden E., Thorgeirsson S. S. In vivo differentiation of rat liver oval cells into hepatocytes. Cancer Res. 1989 Mar 15;49(6):1541–1547. [PubMed] [Google Scholar]
  6. FARBER E. Similarities in the sequence of early histological changes induced in the liver of the rat by ethionine, 2-acetylamino-fluorene, and 3'-methyl-4-dimethylaminoazobenzene. Cancer Res. 1956 Feb;16(2):142–148. [PubMed] [Google Scholar]
  7. Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
  8. Furukawa K., Shimada T., England P., Mochizuki Y., Williams G. M. Enrichment and characterization of clonogenic epithelial cells from adult rat liver and initiation of epithelial cell strains. In Vitro Cell Dev Biol. 1987 May;23(5):339–348. doi: 10.1007/BF02620990. [DOI] [PubMed] [Google Scholar]
  9. GRISHAM J. W. A morphologic study of deoxyribonucleic acid synthesis and cell proliferation in regenerating rat liver; autoradiography with thymidine-H3. Cancer Res. 1962 Aug;22:842–849. [PubMed] [Google Scholar]
  10. GRISHAM J. W., HARTROFT W. S. Morphologic identification by electron microscopy of "oval" cells in experimental hepatic degeneration. Lab Invest. 1961 Mar-Apr;10:317–332. [PubMed] [Google Scholar]
  11. Germain L., Blouin M. J., Marceau N. Biliary epithelial and hepatocytic cell lineage relationships in embryonic rat liver as determined by the differential expression of cytokeratins, alpha-fetoprotein, albumin, and cell surface-exposed components. Cancer Res. 1988 Sep 1;48(17):4909–4918. [PubMed] [Google Scholar]
  12. Germain L., Noël M., Gourdeau H., Marceau N. Promotion of growth and differentiation of rat ductular oval cells in primary culture. Cancer Res. 1988 Jan 15;48(2):368–378. [PubMed] [Google Scholar]
  13. Goyette M., Faris R., Braun L., Hixson D., Fausto N. Expression of hepatocyte and oval cell antigens in hepatocellular carcinomas produced by oncogene-transfected liver epithelial cells. Cancer Res. 1990 Aug 1;50(15):4809–4817. [PubMed] [Google Scholar]
  14. Grisham J. W. Cell types in long-term propagable cultures of rat liver. Ann N Y Acad Sci. 1980;349:128–137. doi: 10.1111/j.1749-6632.1980.tb29521.x. [DOI] [PubMed] [Google Scholar]
  15. Grisham J. W., Tsao M. S., Lee D. C., Earp H. S. Sequential changes in epidermal growth factor receptor/ligand function in cultured rat liver epithelial cells transformed chemically in vitro. Pathobiology. 1990;58(1):3–14. doi: 10.1159/000163560. [DOI] [PubMed] [Google Scholar]
  16. Hiraiwa M., Uda Y. GM1 ganglioside beta-galactosidases from bovine liver. Jpn J Exp Med. 1988 Jun;58(3):129–138. [PubMed] [Google Scholar]
  17. Horan P. K., Slezak S. E. Stable cell membrane labelling. Nature. 1989 Jul 13;340(6229):167–168. doi: 10.1038/340167a0. [DOI] [PubMed] [Google Scholar]
  18. Lee L. W., Raymond V. W., Tsao M. S., Lee D. C., Earp H. S., Grisham J. W. Clonal cosegregation of tumorigenicity with overexpression of c-myc and transforming growth factor alpha genes in chemically transformed rat liver epithelial cells. Cancer Res. 1991 Oct 1;51(19):5238–5244. [PubMed] [Google Scholar]
  19. Lemire J. M., Shiojiri N., Fausto N. Oval cell proliferation and the origin of small hepatocytes in liver injury induced by D-galactosamine. Am J Pathol. 1991 Sep;139(3):535–552. [PMC free article] [PubMed] [Google Scholar]
  20. Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
  21. Marceau N., Blouin M. J., Germain L., Noel M. Role of different epithelial cell types in liver ontogenesis, regeneration and neoplasia. In Vitro Cell Dev Biol. 1989 Apr;25(4):336–341. doi: 10.1007/BF02624596. [DOI] [PubMed] [Google Scholar]
  22. Martínez-Zorzano V. S., Rodríguez Berrocal F. J., Cabezas J. A., Páez de la Cadena M. Comparative studies of two acid beta-galactosidases from rabbit and bovine kidney. Kidney Int. 1989 Jun;35(6):1295–1299. doi: 10.1038/ki.1989.125. [DOI] [PubMed] [Google Scholar]
  23. Ogawa K., Minase T., Onhoe T. Demonstration of glucose 6-phosphatase activity in the oval cells of rat liver and the significance of the oval cells in azo dye carcinogenesis. Cancer Res. 1974 Dec;34(12):3379–3386. [PubMed] [Google Scholar]
  24. Ponder K. P., Gupta S., Leland F., Darlington G., Finegold M., DeMayo J., Ledley F. D., Chowdhury J. R., Woo S. L. Mouse hepatocytes migrate to liver parenchyma and function indefinitely after intrasplenic transplantation. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1217–1221. doi: 10.1073/pnas.88.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Price J., Turner D., Cepko C. Lineage analysis in the vertebrate nervous system by retrovirus-mediated gene transfer. Proc Natl Acad Sci U S A. 1987 Jan;84(1):156–160. doi: 10.1073/pnas.84.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Richter A., Sanford K. K., Evans V. J. Influence of oxygen and culture media on plating efficiency of some mammalian tissue cells. J Natl Cancer Inst. 1972 Dec;49(6):1705–1712. doi: 10.1093/jnci/49.6.1705. [DOI] [PubMed] [Google Scholar]
  27. Rodríguez-Berrocal F. J., Páez de la Cadena M., Cabezas J. A., Pérez-González N. Purification and characterization of acid beta-D-galactosidase from rabbit spleen. J Biochem. 1988 Jul;104(1):66–71. doi: 10.1093/oxfordjournals.jbchem.a122425. [DOI] [PubMed] [Google Scholar]
  28. Sanes J. R., Rubenstein J. L., Nicolas J. F. Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos. EMBO J. 1986 Dec 1;5(12):3133–3142. doi: 10.1002/j.1460-2075.1986.tb04620.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sell S. Is there a liver stem cell? Cancer Res. 1990 Jul 1;50(13):3811–3815. [PubMed] [Google Scholar]
  30. Sell S., Leffert H. L. An evaluation of cellular lineages in the pathogenesis of experimental hepatocellular carcinoma. Hepatology. 1982 Jan-Feb;2(1):77–86. doi: 10.1002/hep.1840020113. [DOI] [PubMed] [Google Scholar]
  31. Sell S., Salman J. Light- and electron-microscopic autoradiographic analysis of proliferating cells during the early stages of chemical hepatocarcinogenesis in the rat induced by feeding N-2-fluorenylacetamide in a choline-deficient diet. Am J Pathol. 1984 Feb;114(2):287–300. [PMC free article] [PubMed] [Google Scholar]
  32. Shiojiri N., Lemire J. M., Fausto N. Cell lineages and oval cell progenitors in rat liver development. Cancer Res. 1991 May 15;51(10):2611–2620. [PubMed] [Google Scholar]
  33. Sirica A. E., Mathis G. A., Sano N., Elmore L. W. Isolation, culture, and transplantation of intrahepatic biliary epithelial cells and oval cells. Pathobiology. 1990;58(1):44–64. doi: 10.1159/000163564. [DOI] [PubMed] [Google Scholar]
  34. Tsao M. S., Grisham J. W., Chou B. B., Smith J. D. Clonal isolation of populations of gamma-glutamyl transpeptidase-positive and -negative cells from rat liver epithelial cells chemically transformed in vitro. Cancer Res. 1985 Oct;45(10):5134–5138. [PubMed] [Google Scholar]
  35. Tsao M. S., Grisham J. W. Hepatocarcinomas, cholangiocarcinomas, and hepatoblastomas produced by chemically transformed cultured rat liver epithelial cells. A light- and electron-microscopic analysis. Am J Pathol. 1987 Apr;127(1):168–181. [PMC free article] [PubMed] [Google Scholar]
  36. Tsao M. S., Grisham J. W., Nelson K. G. Clonal analysis of tumorigenicity and paratumorigenic phenotypes in rat liver epithelial cells chemically transformed in vitro. Cancer Res. 1985 Oct;45(10):5139–5144. [PubMed] [Google Scholar]
  37. Tsao M. S., Grisham J. W., Nelson K. G., Smith J. D. Phenotypic and karyotypic changes induced in cultured rat hepatic epithelial cells that express the "oval" cell phenotype by exposure to N-methyl-N'-nitro-N-nitrosoguanidine. Am J Pathol. 1985 Feb;118(2):306–315. [PMC free article] [PubMed] [Google Scholar]
  38. Tsao M. S., Grisham J. W. Phenotypic modulation during tumorigenesis by clones of transformed rat liver epithelial cells. Cancer Res. 1987 Mar 1;47(5):1282–1286. [PubMed] [Google Scholar]
  39. Tsao M. S., Smith J. D., Nelson K. G., Grisham J. W. A diploid epithelial cell line from normal adult rat liver with phenotypic properties of 'oval' cells. Exp Cell Res. 1984 Sep;154(1):38–52. doi: 10.1016/0014-4827(84)90666-9. [DOI] [PubMed] [Google Scholar]
  40. WILSON J. W., GROAT C. S., LEDUC E. H. HISTOGENESIS OF THE LIVER. Ann N Y Acad Sci. 1963 Dec 30;111:8–24. doi: 10.1111/j.1749-6632.1963.tb36945.x. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES