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Establishment of Tumor Cell Lines: From Primary Tumor Cells to a Tumor Cell Line

  • Katharina Meditz
  • Beate Rinner
Chapter
Part of the Learning Materials in Biosciences book series (LMB)

Abstract

Adequate cell lines and cell systems are the cornerstone of various research fields, with the aim of reflecting the tissue of origin as closely as possible in order to obtain optimal research conditions. Cell lines are defined as continuously growing cell cultures, with unlimited proliferation. During cell line establishment scientists need to be cognizant of numerous contaminants including bacterial, viral and cell-cell interaction along with the issues of Hayflick Limit which can have profound influence over the results, if not successfully avoided. Great patience and attention to detail is needed in order to create a viable specimen.The present chapter will give an overview about handling of tumor tissue, isolation of tumor cells, and finally the establishment of tumor cell lines. Characterization and identification of established cell lines will be shown, on the basis of three examples: clival and sacral chordomas and NRAS-mutated melanoma. Understanding the culture conditions will help to find the perfect requirements for the culturing of each different cell line. Contamination in the cell culture is a touchy topic; detection and elimination of microbial and cell-cell contamination will be discussed in detail. By adhering some golden rules, success of cell line establishment will be guaranteed!

References

  1. 1.
    Aula P, Saksela E. Banding characteristics of paracentric marker constrictions in human chromosomes. Hereditas. 1972;70(2):309–10.CrossRefGoogle Scholar
  2. 2.
    Balkwill FR, Capasso M, Hagemann T. The tumor microenvironment at a glance. J Cell Sci. 2012;125(Pt 23):5591–6.CrossRefGoogle Scholar
  3. 3.
    Boxenberger HJ. Leitfaden für die Zell- und Gewebekultur. Weinheim, Germany: WILEY-VCH Verlag GmbH&Co; 2007.Google Scholar
  4. 4.
    Burdall S, Hanby A, Lansdown R, Speirs V. Breast Cancer cell lines: friend or foe? Breast Cancer Res. 2003;5:89–95.CrossRefGoogle Scholar
  5. 5.
    Clark WH. Tumour progression and the nature of cancer. Br J Cancer. 1991;64(4):631–44. ReviewCrossRefGoogle Scholar
  6. 6.
    Chugh R, Tawbi H, Lucas DR, Biermann JS, Schuetze SM, Baker LH. Chordoma: the nonsarcoma primary bone tumor. Oncologist. 2007;12:1344–50.CrossRefGoogle Scholar
  7. 7.
    Connolly EM, Gaffney E, Reynolds JV. Gastrointestinal stromal tumours. Br J Surg. 2003;90(10):1178–86. ReviewCrossRefGoogle Scholar
  8. 8.
    Cummings BJ, Hodson DI, Bush RS. Chordoma: the results of megavoltage radiation therapy. Int J Radiat Oncol Biol Phys. 1983;9:633–42.CrossRefGoogle Scholar
  9. 9.
    Fletcher CDM, Unni KK, Mertens F, editors. Pathology and genetics of Tumours of soft tissue and bone, World Health Organization classification of Tumours, vol. 5. Lyon: IARC Press; 2002.Google Scholar
  10. 10.
    Freshney RI. Culture of animal cells: a manual of basic technique. 5th ed. Hoboken: Wiley-Liss; 2005.CrossRefGoogle Scholar
  11. 11.
    Gellner V, Tomazic PV, Lohberger B, Meditz K, Heitzer E, Mokry M, Koele W, Leithner A, Liegl-Atzwanger B, Rinner B. Establishment of clival chordoma cell line MUG-CC1 and lymphoblastoid cells as a model for potential new treatment strategies. Sci Rep. 2016;6:24195.CrossRefGoogle Scholar
  12. 12.
    Gey GO, Coffman WD, Kubicek MT. Tissue culture studies of the proliferative capacity of cervical carcinoma and normal epithelium. Cancer Res. 1952;12:364–5.Google Scholar
  13. 13.
    Hallor KH, Staaf J, Jonsson G, Heidenblad M, Vult von Steyern F, Bauer HC, Ijszenga M, Hogendoorn PC, Mandahl N, Szuhai K, Mertens F. Frequent deletion of the CDKN2A locus in chordoma: analysis of chromosomal imbalances using array comparative genomic hybridisation. Br J Cancer. 2008;98:434–42.CrossRefGoogle Scholar
  14. 14.
    Hayflick L, Moorhead PS. The serial cultivation of human diploid cell strains. Exp Cell Res. 1961;25:585–621.CrossRefGoogle Scholar
  15. 15.
    Hussain T, Kotnis A, Sarin R, Mulherkar R. Establishment & characterization of lymphoblastoid cell lines from patients with multiple primary neoplasms in the upper aero-digestive tract & healthy individuals. Indian J Med Res. 2012;135(6):820–9.PubMedPubMedCentralGoogle Scholar
  16. 16.
    Langdon SP. Cancer cell culture methods and protocols. Edingburgh, UK: Human Press., ISSN 1543-1894; 2004.Google Scholar
  17. 17.
    Levine E, Thomas L, McGregor D, Hayflick L, Eagle M. Altered nucleic acid metabolism in human cell cultures infected with mycoplasma. Proc Natl Acad Sci U S A. 1968;60:583–9.CrossRefGoogle Scholar
  18. 18.
    McGarrity GJ, Phillips DM, Vaidya AB. Mycoplasmal infection of lymphocyte cell cultures: infection with M. Salivarium. In Vitro. 1980;16(4):346–56.CrossRefGoogle Scholar
  19. 19.
    Mitra A, Mishra L, Li S. Technologies for deriving primary tumor cells for use in personalized cancer therapy. Trends Biotechnol. 2013;31(6):347–54.CrossRefGoogle Scholar
  20. 20.
    Nelson-Rees WA, Daniles DW, Flandermeyer RR. Cross-contamination of cell culture. Science. 1981;212:446–52.CrossRefGoogle Scholar
  21. 21.
    Nimes R, Sykes G, Cottrill K. Short tandem repeat profiling: part of an overall strategy for reducing the frequency of cell misidentification. In Vitro Cell Dev Biol Anim. 2010;46:811–9.CrossRefGoogle Scholar
  22. 22.
    Perez AG, Kim JH, Gelbard AS, Djordjevic B. Altered incorporation of nucleic acid precursors by mycoplasma-infected mammalia cells in culture. Exp Cell Res. 1972;70:301–10.CrossRefGoogle Scholar
  23. 23.
    Polinger IS. Separation of cell types in embryonic heart cell cultures. Exp Cell Res. 1970;63:78–82.CrossRefGoogle Scholar
  24. 24.
    Rinner B, Froehlich EV, Buerger K, Knausz H, Lohberger B, Scheipl S, Fischer C, Leithner A, Guelly C, Trajanoski S, Szuhai K, Liegl B. Establishment and detailed functional and molecular genetic characterisation of a novel sacral chordoma cell line, MUG-Chor1. Int J Oncol. 2012;40(2):443–51.PubMedGoogle Scholar
  25. 25.
    Rottem S, Barile MF. Beware of mycoplasmas. Trends Biotechnol. 1993;11(4):143–51.CrossRefGoogle Scholar
  26. 26.
    Sklott R. The immortal life of Henrietta Lacks. ISBN9780307888440 Crown; 2010.Google Scholar
  27. 27.
    Stanbridge EJ, Hayflick L, Perkins FT. Modification of amino acid concentrations induced by mycoplasmas in cell culture medium. Nature. 1971;232:242–4.Google Scholar
  28. 28.
    Wise KS, Cassell GH, Action RT. Selective association of murine T lymphoblastoid cell surface alloantigens with mycoplasma hyorhinis. Proc Natl Acad Sci U S A. 1978;75:4479–83.CrossRefGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Division of Biological ResearchMedical University of GrazGrazAustria
  2. 2.Division of Biomedical Research, Core Facility Alternative Biomodels and Preclinical ImagingGrazAustria

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