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. 1980 Oct;101(1):115–126.

Morphologic and histochemical analysis of hepatocytes transplanted into syngeneic hosts.

R L Jirtle, C Biles, G Michalopoulos
PMCID: PMC1903593  PMID: 6108719

Abstract

Suspensions of freshly isolated hepatocytes were prepared by collagenase perfusion of livers of adult Fischer 344 female rats. The cells were injected into the dorsal fascia of 2/3 partially hepatectomized syngeneic hosts (10(6) cells per injection site) and were monitored from 3 days to 3 months after injection. Brown nodules developed at the transplantation site. Histologic examination of the nodules revealed that the hepatocytes were arranged in cords and clusters surrounded by fibrovascular connective tissue. Bile ductules were also seen. Hepatocytes were positive for glucose-6-phosphatase. Staining for gamma-glutamyltranspeptidase showed that the parenchymal hepatocytes were mostly (approximately 95%) negative, whereas bile ductules were positive. These histochemical findings were seen in hepatocytes up to 3 months after transplantation and did not vary with the age of the transplants. Electron-microscopic examination of the transplanted nodules demonstrated that the cells maintained the characteristics of hepatocellular cytoplasmic structure. The relationship between the bile canaliculi and the stromal vessels was found to be similar to the bile canaliculi and hepatic sinusoid polarity seen in the normal liver. Autoradiographic analysis showed that a fraction of the transplanted cells was active in DNA synthesis. This system may become a tool in the study of survival and neoplastic transformation of hepatocytes as a result of exposure to X-irradiation and chemical carcinogens.

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Selected References

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

  1. Bernaert D., Wanson J. C., Drochmans P., Popowski A. Effect of insulin on ultrastructure and glycogenesis in primary cultures of adult rat hepatocytes. J Cell Biol. 1977 Sep;74(3):878–900. doi: 10.1083/jcb.74.3.878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bissell D. M. Study of hepatocyte function in cell culture. Prog Liver Dis. 1976;5:69–82. [PubMed] [Google Scholar]
  4. Cameron R., Sweeney G. D., Jones K., Lee G., Farber E. A relative deficiency of cytochrome P-450 and aryl hydrocarbon [benzo(a)pyrene] hydroxylase in hyperplastic nodules induced by 2-acetylaminofluorene in rat liver. Cancer Res. 1976 Nov;36(11 Pt 1):3888–3893. [PubMed] [Google Scholar]
  5. Clifton K. H., DeMott R. K., Mulcahy R. T., Gould M. N. Thyroid gland formation from inocula of monodispersed cells: early results on quantitation, function, neoplasia and radiation effects. Int J Radiat Oncol Biol Phys. 1978 Nov-Dec;4(11-12):987–990. doi: 10.1016/0360-3016(78)90010-x. [DOI] [PubMed] [Google Scholar]
  6. Decad G. M., Hsieh D. P., Byard J. L. Maintenance of cytochrome P-450 and metabolism of aflatoxin B1 in primary hepatocyte cultures. Biochem Biophys Res Commun. 1977 Sep 9;78(1):279–287. doi: 10.1016/0006-291x(77)91251-7. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Farber E., Parker S., Gruenstein M. The resistance of putative premalignant liver cell populations, hyperplastic nodules, to the acute cytotoxic effects of some hepatocarcinogens. Cancer Res. 1976 Nov;36(11 Pt 1):3879–3887. [PubMed] [Google Scholar]
  9. GRISHAM J. W., LEONG G. F., HOLE B. V. HETEROTOPIC PARTIAL AUTOTRANSPLANTATION OF RAT LIVER: TECHNIC AND DEMONSTRATION OF STRUCTURE AND FUNCTION OF THE GRAFT. Cancer Res. 1964 Sep;24:1474–1495. [PubMed] [Google Scholar]
  10. Gould M. N., Biel W. F., Clifton K. H. Morphological and quantitative studies of gland formation from inocula of monodispersed rat mammary cells. Exp Cell Res. 1977 Jul;107(2):405–416. doi: 10.1016/0014-4827(77)90362-7. [DOI] [PubMed] [Google Scholar]
  11. Guzelian P. S., Bissell D. M., Meyer U. A. Drug metabolism in adult rat hepatocytes in primary monolayer culture. Gastroenterology. 1977 Jun;72(6):1232–1239. [PubMed] [Google Scholar]
  12. LEONG G. F., GRISHAM J. W., HOLE B. V., ALBRIGHT M. L. EFFECT OF PARTIAL HEPATECTOMY ON DNA SYNTHESIS AND MITOSIS IN HETEROTOPIC PARTIAL AUTOGRAFTS OF RAT LIVER. Cancer Res. 1964 Sep;24:1496–1501. [PubMed] [Google Scholar]
  13. Laishes B. A., Farber E. Transfer of viable putative preneoplastic hepatocytes to the livers of syngeneic host rats. J Natl Cancer Inst. 1978 Aug;61(2):507–512. [PubMed] [Google Scholar]
  14. Laishes B. A., Williams G. M. Conditions affecting primary cell cultures of functional adult rat hepatocytes. II. Dexamethasone enhanced longevity and maintenance of morphology. In Vitro. 1976 Dec;12(12):821–832. doi: 10.1007/BF02796367. [DOI] [PubMed] [Google Scholar]
  15. Leffert H. L., Moran T., Boorstein R., Koch K. S. Procarcinogen activation and hormonal control of cell proliferation in differentiated primary adult rat liver cell cultures. Nature. 1977 May 5;267(5606):58–61. doi: 10.1038/267058a0. [DOI] [PubMed] [Google Scholar]
  16. Leffert H., Moran T., Sell S., Skelly H., Ibsen K., Mueller M., Arias I. Growth state-dependent phenotypes of adult hepatocytes in primary monolayer culture. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1834–1838. doi: 10.1073/pnas.75.4.1834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Michalopoulos G., Pitot H. C. Primary culture of parenchymal liver cells on collagen membranes. Morphological and biochemical observations. Exp Cell Res. 1975 Aug;94(1):70–78. doi: 10.1016/0014-4827(75)90532-7. [DOI] [PubMed] [Google Scholar]
  18. Michalopoulos G., Russell F., Biles C. Primary cultures of hepatocytes on human fibroblasts. In Vitro. 1979 Oct;15(10):796–806. doi: 10.1007/BF02618306. [DOI] [PubMed] [Google Scholar]
  19. Michalopoulos G., Sattler C. A., Sattler G. L., Pitot H. C. Cytochrome P-450 induction by phenobarbital and 3-methylcholanthrene in primary cultures of hepatocytes. Science. 1976 Sep 3;193(4256):907–909. doi: 10.1126/science.948753. [DOI] [PubMed] [Google Scholar]
  20. Miyazaki M. Primary culture of adult rat liver cells. II. Cytological and biochemical properties of primary cultured cells. Acta Med Okayama. 1978 Apr;32(1):11–22. [PubMed] [Google Scholar]
  21. Nishizumi M., Albert R. E., Burns F. J., Bilger L. Hepatic cell loss and proliferation induced by N-2-fluorenylacetamide, diethylnitrosamine, and aflatoxin B1 in relation to hepatoma induction. Br J Cancer. 1977 Aug;36(2):192–197. doi: 10.1038/bjc.1977.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Paine A. J., Legg R. F. Apparent lack of correlation between the loss of cytochrome P-450 in hepatic parenchymal cell culture and the stimulation of haem oxygenase activity. Biochem Biophys Res Commun. 1978 Mar 30;81(2):672–679. doi: 10.1016/0006-291x(78)91589-9. [DOI] [PubMed] [Google Scholar]
  23. Rabes H. M., Szymkowiak R. Cell kinetics of hepatocytes during the preneoplastic period of diethylnitrosamine-induced liver carcinogenesis. Cancer Res. 1979 Apr;39(4):1298–1304. [PubMed] [Google Scholar]
  24. Sattler C. A., Michalopoulos G., Sattler G. L., Pitot H. C. Ultrastructure of adult rat hepatocytes cultured on floating collagen membranes. Cancer Res. 1978 Jun;38(6):1539–1549. [PubMed] [Google Scholar]
  25. Savage C. R., Jr, Bonney R. J. Extended expression of differentiated function in primary cultures of adult liver parenchymal cells maintained on nitrocellulose filters. I. Induction of phosphoenolpyruvate carboxykinase and tyrosine aminotransferase. Exp Cell Res. 1978 Jul;114(2):307–315. doi: 10.1016/0014-4827(78)90488-3. [DOI] [PubMed] [Google Scholar]
  26. Sirica A. E., Richards W., Tsukada Y., Sattler C. A., Pitot H. C. Fetal phenotypic expression by adult rat hepatocytes on collagen gel/nylon meshes. Proc Natl Acad Sci U S A. 1979 Jan;76(1):283–287. doi: 10.1073/pnas.76.1.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. WACHSTEIN M., MEISEL E. Histochemistry of hepatic phosphatases of a physiologic pH; with special reference to the demonstration of bile canaliculi. Am J Clin Pathol. 1957 Jan;27(1):13–23. doi: 10.1093/ajcp/27.1.13. [DOI] [PubMed] [Google Scholar]
  28. Williams G. M., Klaiber M., Farber E. Differences in growth of transplants of liver, liver hyperplastic nodules, and hepatocellular carcinomas in the mammary fat pad. Am J Pathol. 1977 Nov;89(2):379–390. [PMC free article] [PubMed] [Google Scholar]
  29. Yager J. D., Jr, Potter V. R. A comparison of the effects of 3'-methyl-4-dimethylaminoazobenzene, 2-methyl-4-dimethylaminoazobenzene, and 2-acetylaminofluorene on rat liver DNA stability and new synthesis. Cancer Res. 1975 May;35(5):1225–1234. [PubMed] [Google Scholar]

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