Immortalisation of Primary Cells



In nature the progression of cells through the cell cycle leading to proliferation of is normally subject to tight regulation by means of the complex and coordinated interaction between cyclins (A-E) and cyclin dependent kinases (for a recent review see Abu-Absi, 2000). ‘Immortalised’ cells escape the normal controls of the cell cycle. Thus, they divide and grow continuously beyond the limits seen in “normal” tissues and primary cells. Historically these continuous cell lines (CCLs) have been primarily isolated from tumours and embryos. In the case of tumours the resulting cell lines may arise from cancer cells that undergo secondary mutations which stabilise or otherwise modify the expression of oncogenes. Viral sequences may also be responsible for the appearance of immortalised cells from cultures of primary cells. Nevertheless, in general where cell lines appear to arise spontaneously from such cultures the mechanism of their apparent replicative immortality is unknown.


Primary Cell Genetically Modify Organism Continuous Cell Line Rapid Cell Proliferation Animal Cell Technology 
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  1. Abu-Absi, N.R. (2000) Cell Cycle Events and Cell Cycle Dependent Processes: Animal Cell technology. in RE Spier (ed), Encyclopedia of Cell technology, Wiley-Interscience, New York, 320–336.Google Scholar
  2. Allen, K.J., Reyes, R., Demmler, K., Mercer. J.F.. Williamson, R. and Whitehead, R.H. (2000) Conditionally immortalised mouse hepatocytes for use in liver gene therapy, J. Gastroenterol. flepatol. 15, 1325–1332.Google Scholar
  3. Bodmar, A.G., Ouellette, M.. Frolkis, M.. Holt, S.E., Chiu, C-P.. Morin, G.B., Harley, C.B., Shay, J.W., Lichtsteiner, S. and Wright, W.E. (1998) Extension of life-span by introduction of telomerase into normal human cells. Science 279. 349–352.CrossRefGoogle Scholar
  4. Brosterhus, H., Brings, S., Leyendeckers, H., Manz, R.A., Miltenyi, S., Radbruch. A., Assenmacher, M., Schmitz, J. (1999) Enrichment and detection of live antigen-specific CD4(+) and CD8(+) T cells based on cytokine secretion, Eur. J. Immunol. 29. 4053–4039.Google Scholar
  5. Bryan, T.M. and Reddel, R.R. (1997) Telomere dynamics and telomerase activity in in vitro immortalised human cells. Eur.J.Can. 33, 767–773.CrossRefGoogle Scholar
  6. Darnbrough, C., Slater, S., Vass,M. and MacDonald, C. (1992) Immortalization of murine primary spleen cells by v-myc. v-ras, and v-rat; Exp. Cell Res. 201, 273–283.Google Scholar
  7. Doyle, A. and Griffiths, J.B. (2000) Cell and Tissue Culture for Medical Research, John Wiley & Sons, Chichester, UK.Google Scholar
  8. Erbacher, P. Roche, A.C., Monsigny, M. and Midoux, P. (1995) Glycosylated polylysine/DNA complexes: gene transfer efficiencies in relation with the size and sugar substitution level of glycosylated polylysines and with the plasmid size. Bioconjugate Chem. 6, 401–410.CrossRefGoogle Scholar
  9. Freshney, I.R. (1994) Culture of Animal Cells: A Manual of Basic Technique ( Third Edition) Wiley-Liss, New York.Google Scholar
  10. Fry, J. and Bridges, J.W. (1979) The effect of phenobarbitone on adult rat liver cells and primary cell lines, Lett. 4, 295–301.Google Scholar
  11. Gingrich, J.R. and Roder, J. (1998) Inducible gene expression in the nervous system of transgenic mice, Ann. Rev. Neurosci. 21, 377–405.PubMedCrossRefGoogle Scholar
  12. Hallauer, C., Kronauer, G. and Siegl, G. (1971) Parvovirus contaminants of permanent human cell lines I virus isolation from 1960–1970. Arch. Gesamte l’irusforsch. 35. 80–90.CrossRefGoogle Scholar
  13. Hawley. R.G., Hawley, T.S., Fong A.Z.C., Quinto. C., Collins. M.. Leonard, J.P. and Goldman, S.J. (1996) Thrombopoietic potential and serial repopulating ability of murine hematopoietic stem cells constitutively expressing interleukin 1 I. Proc. Natl. Acad. Sci. 93, 10297–10302.Google Scholar
  14. Herwig, S. and Strauss, M. (1997) The retinoblastoma protein: a master regulator of cell cycle, differentiation and agpoptosis, Eur. J. Biochem. 246. 581–601.PubMedCrossRefGoogle Scholar
  15. Holley, M.C. and Lawlor, P.W. (1997) Production of conditionally immortalised cell lines from a transgenic mouse, Audiol. Neurootol. 2, 25–35.PubMedCrossRefGoogle Scholar
  16. Hug, P. and Sleight, R.G. (1991) Liposomes for the transformation of eukaryotic cells. Biochem. Biophys. Acta 1097, 1–17.PubMedCrossRefGoogle Scholar
  17. Jordan, M., Shallhorn, A. and Wurm. F.M. (1996) Transfecting mammalian cells: optimization of critical parameters affecting calcium-phosphate precipitate formation.,Nucleic Acids Res. 24, 596–601Google Scholar
  18. Kao, W-Y and Prockop, D.I. (1977) Proline analogue removes fibroblasts from cultured mixed cell populations. Nature 266, 63–64.PubMedCrossRefGoogle Scholar
  19. Lee, R.J. and Huang, L. (1997) Lipidic vector systems for gene transfer. Crit. Rev. in Therapeutic Drag Carrier Systems 14 173–206.Google Scholar
  20. Littlewood, T.D., Hancock, D.C., Danielian, P.S.. Parker, M.G. and Evan, G. (1995) A modified receptor ligand-binding domain as an improved switch for the regulation of heterologous proteins, Nucleic Acids Res. 23. 1686–1690.Google Scholar
  21. MacDonald, C. (1998) Safety aspects of genetic modification procedures, in Stacey. GN, Doyle, A, Hambleton, P (eds.) Safety Considerations in Cell and Tissue Culture. Kluwer Academic Publishers, Dordrecht, pp 189–204.CrossRefGoogle Scholar
  22. Maurer, N., Mori. A., Palmer. L., Monck, M.A., Mok. K.W.C., Mui, B., Akhong, Q.F. and Cullis, P.R. (1999) Lipid-based systems for the intracellular delivery of genetic drugs. Mol. Memb. Biol. 16, 129140.Google Scholar
  23. Mayne, L.V., Priestley, A., James, M.R. and Burke, J.F. (1986) Efficient immortalization and morphological transformation of human fibroblasts by transfection with SV40 DNA linked to a dominant marker, Exp. Cell Res. 162, 530–538.PubMedCrossRefGoogle Scholar
  24. McLean J.S. (1999) Immortalisation strategies for mammalian cells, in Jenkins (ed), Methods in Biotechnology, Vol. 8 Animal Cell Biotechnology,Humana Press Inc, Totowa, NJ, pp.61–72.Google Scholar
  25. Morales, C.P., Holt, S.E., Ouellette, M., Kaur, K..I., Van, Y., Wilson, K.S., White, M.A., Wright, W.A. and Shay, J.W. (1999) Absence of cancer associated changes in human fibroblasts immortalised with telomerase. Nature Genetics 21, 115–118.PubMedCrossRefGoogle Scholar
  26. Parkinson, E.K., Newbold, R.F. and Keith, W.N. (1997) The genetic basis of human keratinocyte immortalisation in squamous cell carcinoma development: the role of telomerase reactivation, Eur. J. Can. 33. 727–734.CrossRefGoogle Scholar
  27. Popovic, M., Lange-Wantzin, G., Sarin, P.S., Mann. D. and Gallo, R.C. (1983) Transformation of umbilical cord blood T cells by human T-cell leukaemia/lymphoma virus, Proc. Natl. Acad. Sci. 80, 5402–5406.Google Scholar
  28. Reddel, R.R., Ke. Y., Gerwin. B.I., McMenamin, M.G., Lechner, J.F.. Su, R.T., Brash, D.E., Park, J.B., Rhim, I.S. and Harris. C.C. (1988) Transformation of human bronchial epithelial cells by infection with SV40 or adenovirus-12 SV40 hybrid virus. or transfection by strontium phosphate coprecipitation with a plasmid containing SV40 early region genes. Cancer Res. 48, 1904–1909.PubMedGoogle Scholar
  29. Sandig, V.. Lieber, A. and Strauss, M. (1997) lectors for gene transfer and expression in animal cells, in Mammalian Cell Biotechnology in Protein Product, Walter de Gruyter, Berlin/New York. pp. 65–85.Google Scholar
  30. Schlokat, U., Himmelspach, M., Falkner, F.G., and Dourer, F. (1997) Permanent gene expression in mammalian cells, in Mammalian Cell Biotechnology in Protein Production, Walter de Gruyter, Berlin/New York, pp. 37–52.Google Scholar
  31. Schwarzenberger, P., P., Spence, S.E., Gooya, J.M., Michiel, D.. Curiel, D.T., Ruscetti, F.W. and Keller, J.R. (1996) Targeted gene transfer to human haematopoietic progenitor cell lines through the c-kit receptor. Blood 87, 472–478.PubMedGoogle Scholar
  32. Scott, D..M., MacDonald, C.. Brzeski, H. and Kinne, R. (1986) Maintenance and expression of differentiated function of kidney cells following transformation by SV40 early region DNA. Exp. Cell Res. 166, 391–398.Google Scholar
  33. Shirata, S., Truyak, K., Mori, T., Seki, K., Ohashi, H.. Tachibana, H. and Murakami. H. (1991) Genetic enhancement of protein productivity of animal cells by oncogene, in K. Sasaki and K. Ikura (eds), Animal Cell Culture and Production of Biologicals. Kluwer Academic Publishers, Dordrecht, pp 259–266.CrossRefGoogle Scholar
  34. Shigekawa, K. and Dower, W.J. (1988) Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. Biotechniques 6, 742–751.PubMedGoogle Scholar
  35. Stacey, G.N. and Doyle, A. (2000) Cell Banks: A Service to Animal Cell Technology, in R.E. Spier Encyclopedia of Cell Technology., Wiley-lnterscience, New York, pp 293–320.Google Scholar
  36. Stacey, G.N., Masters, J.R., MacLeod, R.A.F., Drexler, H. and Freshney. I.R. (2000) Cell contamination leads to inaccurate darta: we must take action now. Nature 403, 356.PubMedCrossRefGoogle Scholar
  37. Tamai, T., Sato, N., Kimura, S.. Shirahata, S. and Murakami, H. (1992) Immortalisation of flatfish (Paralichthys olivaccus) leukocytes by oncogene transfection, in R.E. Spier, J.B. Griffiths and C. MacDonald (eds.), Animal Cell Biotechnology: Developments. Processes and Products, Butterworth-Heinemann, Oxford, UK, pp. 29–31.Google Scholar
  38. Twyman, R.M. and Whitelaw, B (2000) Genetic Engineering: Animal Cell Technology, in R.E.Spier (ed.), Encyclopedia of Cell technology, Wiley-Interscience, New York, pp. 737–819.Google Scholar
  39. United Kingdom Co-ordinating Committee on Cancer Research Ad Hoc Working Party (1999) UKCCCR Guidelines for the Use of Cell Lines in Cancer Research. UKCCCR, PO Box 123, Lincoln’s Inn Fields, London.Google Scholar
  40. Vaziri, H.F. and Bechimol, S. (1998) Reconstitution of telomerase activityin normal human cells leads to elongation of telomeres and extended replicative life span. Curr. Biol. 8, 279–282.PubMedCrossRefGoogle Scholar
  41. Vaziri, H.F. and Bechimol, S. (1999) Alternative pathways for the extention of cellular life span: inactivation of p53/Rb and expression of telomerase. Oncogene 18, 7676–7680.PubMedCrossRefGoogle Scholar
  42. Werner, A., Duvar, S., Muthing J., Buntemeyer, H., Kahmann, U., Lunsdorf. H. and Lehmann, J. (1999) Cultivation and characterisation of a new immortalised human hepatocyte cell line, Hep Z, for use in an artificial liver support system, Ann. NY Acad. Sci. 875, 364–368.Google Scholar
  43. Wu, G.Y. and Wu, C.H. (1987) Receptor mediated in vitro gene transformation by a soluble DNA carrier system. J. Biol. Chem. 262, 4429–4432.PubMedGoogle Scholar
  44. Yang, J., C hang, E., Cherry, A.M., Bangs, C.D., Oei, Y., Bodnar, A., Bronstein, A., Chui, C-P. and Herron, G.S. (1999) Human endothlial cell life extension by telomerase expression. J. Biol. Chem. 274, 26141–26148.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2001

Authors and Affiliations

  1. 1.NIBSCSouth Mimms, HertsUK
  2. 2.University of PaisleyGlasgowScotland, UK

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