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Generation of Human Cell Lines Using Lentiviral-Mediated Genetic Engineering

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Epithelial Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 945))

Abstract

Even now, most human cell lines used in research are derived from tumor cells. They are still widely used because they grow well in vitro and so far have helped answering several basic biological questions. However, as modern biology moves into more sophisticated areas, scientists now need human cell lines closer to normal primary cells and further from transformed cancerous cells. The recent identification of cellular genes involved in cell cycling and senescence, together with the development of molecular tools capable of cleanly integrating transgenes into the genome of target cells, have moved the frontier of genetic engineering. In this chapter, we present a detailed hands-on protocol, based on lentivirus-derived vectors and a combination of two native cellular genes that has proven very efficient in generating immortal cell lines from several human primary cells, while preserving most of their original properties.

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Acknowledgment

This work is supported by the Chicago Diabetes Project (http://www.chicagodiabetesproject.org).

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Authors and Affiliations

  1. Department of Neurosciences, Geneva School of Medicine (CMU), Geneva, Switzerland

    Patrick Salmon

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  1. Patrick Salmon
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Correspondence to Patrick Salmon .

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Editors and Affiliations

  1. Cystic Fibrosis/Pulmonary Research Ctr., University of North Carolina, Thurston Bowles Building 4011, Chapel Hill, 27599, North Carolina, USA

    Scott H. Randell

  2. Cystic Fibrosis/Pulmonary Research Ctr., The University of North Carolina, Thurston-Bowles Building 4011, Chapel Hill, 27599, North Carolina, USA

    M. Leslie Fulcher

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Salmon, P. (2012). Generation of Human Cell Lines Using Lentiviral-Mediated Genetic Engineering. In: Randell, S., Fulcher, M. (eds) Epithelial Cell Culture Protocols. Methods in Molecular Biology, vol 945. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-125-7_25

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  • DOI: https://doi.org/10.1007/978-1-62703-125-7_25

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-124-0

  • Online ISBN: 978-1-62703-125-7

  • eBook Packages: Springer Protocols

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