Skip to main content
SpringerLink
Log in

Conditions for growth of hepatocytes in culture

  • Chapter
The Hepatocyte Review

Abstract

Investigators have a variety of reasons for studying the factors that determine the ability of hepatocytes to grow, and for pursuing some of these studies utilising the relatively defined and easily-manipulated conditions of cell culture. There are intriguing intellectual and technical challenges in dissecting out the factors that determine patterns of hepatocyte growth, death and differentiation — the cell lineage relationships, the relevant humoral and nutritional factors and the interactions with extracellular matrix and with non-hepatocyte cell types present in the liver. Some notable issues in biology and medicine underlie efforts to address these challenges. These include a need to understand the ability of generally healthy liver to rapidly replace lost tissue via liver regeneration; to understand normal patterns of hepatocyte growth and their control and the perturbations of these processes relevant to liver carcinogenesis; and to understand factors relevant to regeneration and repair when liver is damaged by toxins (including alcohol) or by virus infection. In addition, a better understanding of hepatocyte growth (and its relationship to cell differentiation or death) could assist efforts to replace defective liver function either with transplanted hepatocytes or with extracorporeal bioartificial livers. For example, ability to manipulate growth in vitro could increase options for generating normal or genetically-modified hepatocytes for transplantation or for developing bioartificial liver devices with sufficient cell mass, appropriate tissue architecture and extended function necessary for successful application in replacing the function of failing human livers.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Isolated hepatocytes: Preparation, properties and applications. Berry MN, Edwards AM and Barritt GJ, Eds. Amsterdam: Elsevier, 1991.

    Google Scholar 

  2. Seglen PO. Preparation of isolated liver cells. Methods Cell Biol 1976; 13: 29–83.

    Article  PubMed  CAS  Google Scholar 

  3. Overturf K, Al-Dhalimy M, Ou C-N, Finegold M and Grompe M. Serial transplantation reveals the stem-cell-like regenerative potential of adult mouse hepatocytes. Amer J Pathol 1997; 151: 1273–1280.

    CAS  Google Scholar 

  4. Bucher NLR and Farmer SR. Liver regeneration following partial hepatectomy: genes and metabolism. In: Liver growth and repair. Strain AJ and Diehl AM, Eds. London: Chapman Hall, 1998, pp 3–27.

    Chapter  Google Scholar 

  5. Fausto N and Webber EM. Liver regeneration. In: The Liver: Biology and Pathobiology ( 3rd ed. ). Arias IM, Boyer.TL, Fausto N, Jakoby WB, Schachter DA and Shafritz DA, Eds. New York: Raven Press, 1994, pp 1059–1084.

    Google Scholar 

  6. Mossin L, Blankson H, Huitfeldt H and Seglen PO. Ploidy-dependent growth and binucleation in cultured rat hepatocytes. Exp Cell Res 1994; 214: 551–560.

    Article  PubMed  CAS  Google Scholar 

  7. Seglen PO. DNA ploidy and autophagic protein degradation as determinants of hepatocellular growth and survival. Cell Biol Toxicol 1997; 13: 301–315.

    Article  PubMed  CAS  Google Scholar 

  8. Rajvanshi P, Liu D, Ott M, Gagandeep S, Schilsky ML and Gupta S. Ploidy-based fractionation of rat hepatocyte subpopulations with varying metabolic potential, proliferative capacity and retroviral gene transfer efficiency. Exp Cell Res 1998; 244: 405–419.

    Article  PubMed  CAS  Google Scholar 

  9. Roth GS. Age changes in signal transduction and gene expression. Mech Ageing Dev 1997; 98: 231–238.

    Article  PubMed  CAS  Google Scholar 

  10. Sawada N, Kojima T, Obata H, Saitoh M, Isomura K, Kokai Y, Satoh M and Mori M. P21 waf-l/cip-1/sdi-1 is expressed at G1 phase in primary culture of hepatocytes from old rats, presumably preventing the cells from entering the S-phase of the cell cycle. Biochem Biophys Res Commun 1996; 228: 819–824.

    Article  PubMed  CAS  Google Scholar 

  11. Coleman WB and Grisham JW. Epithelial stem-like cells of the rodent liver. In: Liver growth and repair. Strain AJ and Diehl AM, Eds. London: Chapman Hall, 1998, pp 50–99.

    Chapter  Google Scholar 

  12. Petersen BE, Bowen WC, Patrene KD, Mars WM, Sullivan AK et al. Bone marrow as a potential source of hepatic oval cells. Science 1999; 284: 1168–1170.

    Article  PubMed  CAS  Google Scholar 

  13. Enat R, Jefferson DM, Nelson RM, Gatmaitan Z, Leinward LA and Reid LM. Hepatocyte proliferation in vitro: its dependence on the use of serum-free, hormonally defined medium and substrata of extracellular matrix. Proc Natl Acad Sci USA 1984; 81: 1411–1415.

    Article  PubMed  CAS  Google Scholar 

  14. Shimaoka S, Nakamura T and Ichihara A. Stimulation of growth of primary cultured adult rat hepatocytes without growth factors by coculture with non-parenchymal liver cells. Exp Cell Res 1987; 172: 228–242.

    Article  PubMed  CAS  Google Scholar 

  15. Maher JJ. The extracellular matrix in liver regeneration. In: Liver Growth and Repair. Strain AJ and Diehl AM, Eds. London: Chapman Hall, 1998, 451–464.

    Chapter  Google Scholar 

  16. Kim T-H, Mars WM, Stolz DB, Petersen BE and Michalopoulos GK. Extracellular matrix remodelling at the early stages of liver regeneration in the rat. Hepatology 1997; 26: 896–904.

    Article  PubMed  CAS  Google Scholar 

  17. Fladmark KE, Gjertsen BT, Molven A, Mellgren G, Vintermyr OK and Doskeland SO. Gap junctions and growth control in liver regeneration and in isolated rat hepatocytes. Hepatology 1997; 25: 847–855.

    Article  PubMed  CAS  Google Scholar 

  18. Liu ML, Mars WM, Zarnegar R and Michalopoulos GK. Collagenase pretreatment and the mitogenic effects of hepatocyte growth factor and transforming growth factor-a in adult rat liver. Hepatology 1994; 19: 1521–1527.

    Article  PubMed  CAS  Google Scholar 

  19. Tygstrup N, Jensen SA, Krog B, Dietranjelo A and Shafritz DA. Expression of messenger RNA for liver functions following 70% and 90% hepatectomy. J Hepatol 1996; 25: 72–78.

    Article  PubMed  CAS  Google Scholar 

  20. Taub R. Liver regeneration 4: transcriptional control of liver regeneration. FASEB J 1996; 10: 413–427.

    PubMed  CAS  Google Scholar 

  21. Rana BD, Mischoulon Y, Xie Y, Bucher NL and Farmer SR. Cell-extracellular matrix interactions can regulate the switch between growth and differentiation in rat hepatocytes: reciprocal expression of C/EBPa and immediate-early growth response transcription factors. Mol Cell Biol 1994; 14: 5858–5869.

    Article  PubMed  CAS  Google Scholar 

  22. Mitaka T, Mizuguchi T, Sato F, Mochizuchi C and Mochizuki Y. Growth and maturation of small hepatocytes. J Gastroent Hepatol 1998; 13 (suppl): S70 - S77.

    Google Scholar 

  23. Block GD, Locker J, Bowen WC, Petersen BE, Katyal S et al. Population expansion, clonal growth and specific differentiation patterns in primary cultures of hepatocytes induced by HGF/SF, EGF and TGF-a in a chemically defined (HGM) medium. J Cell Biol 1996; 132: 1133–1149.

    Article  PubMed  CAS  Google Scholar 

  24. Cable EE and Isom HC. Exposure of primary rat hepatocytes in long-term DMSO culture to selected transition metals induces hepatocyte proliferation and formation of duct-like structures. Hepatology 1997; 26: 1444–1457.

    Article  PubMed  CAS  Google Scholar 

  25. Tateno C and Yoshizato K. Growth and differentiation of adult rat hepatocytes regulated by the interaction between parenchymal and non-parenchymal liver cells. J Gastroent Hepatol 1998; 13 (suppl): S83 - S92.

    Google Scholar 

  26. Mitaka T, Sattler CA, Sattler GL, Sargent LM and Pitot HC. Multiple cell cycles occur in rat hepatocytes cultured in the presence of nicotinamide and epidermal growth factor. Hepatology 1991; 13: 21–30.

    Article  PubMed  CAS  Google Scholar 

  27. Li Y, Sattler GL and Pitot HC. The effect of amino acid composition of serum-free medium on DNA synthesis in primary hepatocyte cultures in the presence of epidermal growth factor. In Vitro Cell Dev Biol Anim 1995; 31: 867–870.

    Article  CAS  Google Scholar 

  28. Mitaka T, Mikami M, Sattler GL, Pitot HC and Mochizuki Y. Small cell colonies appear in the primary culture of adult rat hepatocytes in the presence of nicotinamide and epidermal growth factor. Hepatology 1992; 16: 440–447.

    Article  PubMed  CAS  Google Scholar 

  29. Tateno C and Yoshizato K. Long-term cultivation of adult rat hepatocytes that undergo multiple cell divisions and express normal parenchymal phenotypes. Amer J Pathol 1996; 148: 383–392.

    CAS  Google Scholar 

  30. Tateno C and Yoshizato K. Growth and differentiatioon in culture of clonigenic hepatocytes that express both phenotypes of hepatocytes and biliary epithelial cells. Amer J Pathol 1996; 149: 1593–1605.

    CAS  Google Scholar 

  31. Mitaka T, Kojima T, Mizuguchi T and Mochizuki Y. Subculture of proliferating adult rat hepatocytes in medium supplemented with nicotinamide and EGF. In Vitro Cell Dev Biol Anim 1996; 32: 469–477.

    Article  CAS  Google Scholar 

  32. Mitaka T, Norioka K and Mochizuki Y. Redifferentiation of proliferated rat hepatocytes cultured in L-15 medium supplemented with EGF and DMSO. In Vitro Cell Dev Biol Anim 1993; 29A: 714–722.

    Article  Google Scholar 

  33. Kost DP and Michalopoulos GK. Effect of 2% dimethylsulfoxide on the mitogenic properties of epidermal growth factor and hepatocyte growth factor in primary hepatocyte culture. J Cell Physiol 1991; 147: 274–280.

    Article  PubMed  CAS  Google Scholar 

  34. Chan K, Kost DP and Michalopoulos GK. Multiple sequential periods of DNA synthesis and quiescence in primary hepatocyte cultures maintained on the DMSOEGF on/off protocol. J Cell Physiol 1989; 141: 584–590.

    Article  PubMed  CAS  Google Scholar 

  35. Bour ES, Ward LK, Cornman GA and Isom HC. Tumor necrosis factor-a-induced apoptosis in hepatocytes in long-term culture. Amer J Pathol 1996; 148: 485–495.

    CAS  Google Scholar 

  36. Michalopoulos GK, Bowen W, Nussler AK, Becich MJ and Howard TA. Comparative analysis of mitogenic and morphogenic effects of HGF/SF and EGF on rat and human hepatocytes maintained in collagen gels. J Cell Physiol 1993; 156: 443452.

    Google Scholar 

  37. Sakata H, Takayama H, Sharp R, Rubin JS, Merlino G and La Rochelle WJ. Hepatocyte growth factor/scatter factor overexpression induces growth, abnormal development and tumor formation in transgenic mouse livers. Cell Growth Differ 1996; 7: 1513–1523.

    PubMed  CAS  Google Scholar 

  38. Hansen LK, Mooney DJ, Vacanti JP and Ingber DE. Integrin binding and cell spreading on extracellular matrix act at different points in the cell cycle to promote hepatocyte growth. Mol Biol Cell 1994; 5: 967–975.

    PubMed  CAS  Google Scholar 

  39. Kim TH, Bowen WC, Stolz DB, Runge D, Mars WM and Michalopoulos GK. Differential expression and distribution of focal adhesion and cell adhesion molecules in rat hepatocyte differentiation. Exp Cell Res 1998; 244: 93–104.

    Article  PubMed  CAS  Google Scholar 

  40. Webber EM, Bruix J, Pierce RH and Fausto N. Tumor necrosis factor primes hepatocytes for DNA replication in the rat. Hepatology 1998; 28: 1226–1234.

    Article  PubMed  CAS  Google Scholar 

  41. Watanable Y, Osaki H and Akaike T. TNF-a bifunctionally induces proliferation in primary hepatocytes: role of cell anchorage and spreading. J Immunol 1997; 159: 4840–4847.

    Google Scholar 

  42. Rolfe M, James NH and Roberts RA. Tumor necrosis factor alpha (TNF-a) suppresses apoptosis and induces DNA synthesis in rodent hepatocytes: a mediator of the hepatocarcinogenicity of peroxisome proliferators? Carcinogenesis 1997; 18: 2277–2280.

    Article  PubMed  CAS  Google Scholar 

  43. Fausto N. Mechanisms of hepatocyte proliferation and apoptosis induced by tumor necrosis factor. 8th Biennial International Congress on Liver Development, Gene Regulation and Disease, Orvietto, Italy 1999: 14.

    Google Scholar 

  44. Chevalier S, MacDonald N and Roberts RA. Nafenopin-induced S-phase is independent of both TNF-a priming activity and EGF receptor tyrosine kinase activity. 8th Biennial International Congress on Liver Development, Gene Regulation and Disease, Orvietto, Italy, 1999: 39.

    Google Scholar 

  45. Korver W, Schilham MW, Moerer P, van den Hoff MJ, Dam K, Lamers WH, Medema RH and Clevers H. Uncoupling of S-phase and mitosis in cardiomyocytes and hepatocytes lacking the winged-helix transcription factor trident. Current Biol 1998; 8: 1327–1330.

    Article  CAS  Google Scholar 

  46. Michalopoulos GK, Bowen WC, Zajak VF, Beer-Stolz D, Watkins S, Kostrubsky V and Strom SC. Morphogenetic events in mixed cultures of rat hepatocytes and non-parenchymal cells maintained in biological matrices in the presence of hepatocyte growth factor and epidermal growth factor. Hepatology 1999; 29: 90–100.

    Article  PubMed  CAS  Google Scholar 

  47. Mitaka T, Sato F, Mizuguchi T, Yokono T and Mochizuchi Y. Construction of hepatic organoid by rat small hepatocytes and hepatic non-parenchymal cells. Hepatology 1999; 29: 111–125.

    Article  PubMed  CAS  Google Scholar 

  48. Takeshita K, Bowen WC and Michalopoulos GK. Three-dimensional culture of hepatocytes in a continuously-flowing medium. In Vitro Cell Dev Biol Anim 1998; 34: 482–485.

    Article  CAS  Google Scholar 

  49. Maher JJ, Bissell DM, Friedman SL and Roll FJ. Collagen measured in primary cultures of normal rat hepatocytes derives from lipocytes within the monolayer. J Clin Invest 1988; 82: 450–459.

    Article  PubMed  CAS  Google Scholar 

  50. Simpson GEC and Finckh ES. The pattern of regeneration of rat liver after repeated partial hepatectomies. J Pathol Bacteriol 1963; 86: 361–370.

    Article  PubMed  CAS  Google Scholar 

  51. Wu JC, Merlino G, Cveklova K, Mosinger B and Fausto N. Autonomous growth in serum-free medium and production of hepatocellular carcinomas by differentiated hepatocyte lines that over-express transforming growth factor a. Cancer Res 1994; 54: 5964–5973.

    PubMed  CAS  Google Scholar 

  52. Hino H, Tateno C, Sato H, Yamasaki C, Katayama S et al. A long-term culture of human hepatocytes which show a high growth potential and express their differentiated phenotypes. Biochem Biophys Res Comm 1999; 256: 184–191.

    Article  PubMed  CAS  Google Scholar 

  53. Runge DM, Runge D, Dorko K, Pisarov LA, Leckel K et al. Epidermal growth factor-and hepatocyte growth factor-receptor activity in serum-free cultures of human hepatocytes. J Hepatol 1999; 30: 265–274.

    Article  PubMed  CAS  Google Scholar 

  54. Mitaka T, Norioka K, Nakamura T and Mochizuki Y. Effects of mitogens and comitogens on the formation of small-cell colonies in primary cultures of rat hepatocytes. J Cell Physiol 1993; 157: 461–468.

    Article  PubMed  CAS  Google Scholar 

  55. Lazaro CA, Rhim JA, Yamada Y and Fausto N. Generation of hepatocytes from oval cell precursors in culture. Cancer Res 1998; 58: 5514–5522.

    PubMed  CAS  Google Scholar 

  56. Runge D, Runge DM, Drenning SD, Bowen WC, Grandis JR and Michalopoulos GK. Growth and differentiation of rat hepatocytes: changes in transcription factors HNF-3, HNF-4, STAT-3 and STAT-5. Biochem Biophys Res Commun 1998; 250: 762–768.

    Article  PubMed  CAS  Google Scholar 

  57. Runge D, Runge DM, Daskalakis N, Lubecki KA, Bowen WC and Michalopoulos GK. Matrix-mediated changes in the expression of HNF4a isoforms and in DNA-binding activity of ARP-1 in primary cultures of rat hepatocytes. Biochem Biophys Res Commun 1999; 259: 651–655.

    Article  PubMed  CAS  Google Scholar 

  58. Michalopoulos GK. Liver regeneration: molecular mechanisms of growth control. FASEB J 1990; 4: 176–187.

    PubMed  CAS  Google Scholar 

  59. Cruise JL, Houck KA and Michalopoulos GK. Induction of DNA synthesis in cultured hepatocytes through stimulation of the a-1 adrenoreceptor by norepinephrine. Science 1985; 15: 749–751.

    Google Scholar 

  60. Ni N and Yager JD. Comitogenic effects of estrogens on DNA synthesis induced by various growth factors in cultured female rat hepatocytes. Hepatology 1994; 19: 183192.

    Google Scholar 

  61. Yusof YA and Edwards AM. Stimulation of DNA synthesis in primary rat hepatocyte cultures by liver tumor promoters: interactions with other growth factors. Carcinogenesis 1990; 11: 761–770.

    Article  PubMed  CAS  Google Scholar 

  62. Fausto N, Laird AD and Webber EM. Liver regeneration 2. Role of growth factors and cytokinase in liver regeneration. FASEB J 1995; 9: 1527–1536.

    PubMed  CAS  Google Scholar 

  63. Nishikawa Y, Wang M and Carr BI. Changes in TGF-ß receptors of rat hepatocytes during primary culture and liver regeneration: increased expression of TGF-(3 receptors associated with increased sensitivity to TGF-13-mediated growth inhibition. J Cell Physiol 1998; 176: 612–623.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Medical Biochemistry, Flinders University, GPO Box 2100, Adelaide, South Australia, Australia, 5001

    Anthony M. Edwards

  2. Department of Pathology, University of Pittsburgh Medical School, Pittsburgh, Pennsylvania, USA, 1526

    George K. Michalopoulos

Authors
  1. Anthony M. Edwards
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. George K. Michalopoulos
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Michael N. Berry Anthony M. Edwards

Rights and permissions

Reprints and permissions

Copyright information

© 2000 Springer Science+Business Media Dordrecht

About this chapter

Cite this chapter

Edwards, A.M., Michalopoulos, G.K. (2000). Conditions for growth of hepatocytes in culture. In: Berry, M.N., Edwards, A.M. (eds) The Hepatocyte Review. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-3345-8_7

Download citation

  • DOI: https://doi.org/10.1007/978-94-017-3345-8_7

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-90-481-5402-9

  • Online ISBN: 978-94-017-3345-8

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation