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Genetic Modification of Human Primary Keratinocytes by Lentiviral Vectors

  • Ana Freije
  • Natalia Sanz-Gómez
  • Alberto GandarillasEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 2109)

Abstract

Keratinocytes are hard to transfect. Viral vectors are a good alternative to genetically modify primary keratinocytes. A classical method is the use of retroviral vectors by co-culture of keratinocytes with virus-producer cells. This method is efficient in high-calcium conditions with feeder cells. However, sometimes co-culture is not possible and is more laborious as producer cells need to be replaced by feeder cells. Our solution is the use of lentiviral vectors, far more efficient as supernatant on keratinocytes. In this chapter we describe improved detailed protocols for stable genetic modification of human primary keratinocytes of the skin or head and neck, in both low- and high-calcium conditions by lentiviral vectors.

Keywords

Keratinocytes Viral vectors Feeder cells Calcium conditions Lentiviral vectors Genetic modification 

Notes

Acknowledgments

This work was funded by Instituto de Salud Carlos III/FEDER (AG; Spain), grants PI14/00900 and PI17/01307. NSG is recipient of a predoctoral scholarship from Universidad de Cantabria/IDIVAL (Spain).

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Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Ana Freije
    • 1
  • Natalia Sanz-Gómez
    • 1
  • Alberto Gandarillas
    • 1
    • 2
    Email author
  1. 1.Cell Cycle, Stem Cell Fate and Cancer LaboratoryInstitute for Research Marqués de Valdecilla (IDIVAL)SantanderSpain
  2. 2.INSERM, Languedoc-RoussillonMontpellierFrance

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