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Stable Genetic Modification of Mesenchymal Stromal Cells Using Lentiviral Vectors

  • Francisco MartínEmail author
  • María Tristán-Manzano
  • Noelia Maldonado-Pérez
  • Sabina Sánchez-Hernández
  • Karim Benabdellah
  • Marién Cobo
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1937)

Abstract

Mesenchymal stromal cell (MSC) therapy has produced very promising results for multiple diseases in animal models, with over 780 clinical trials on going or completed. However, most of the human clinical trials have not been as successful as trials using preclinical models. To improve the therapeutic potential of MSCs, different research groups have used gene transfer vectors to express factors involved in migration, survival, differentiation, and immunomodulation. The ideal gene transfer vector for most applications should achieve long-term, stable (constitutive or inducible) transgene expression in MSCs and their progeny. Given their efficiency and low impact on transduced cells, lentiviral vectors (LVs) are the vectors of choice. In this chapter we will describe a detailed protocol for the generation of genetically modified MSCs using lentiviral vectors (LVs). Although this protocol has been optimized for MSC lentiviral transduction, it can be easily adapted to other stem cells by changing culture conditions while maintaining volumes and incubation times.

Key words

Mesenchymal stromal cells MSCs ASCs Lentiviral vectors Constitutive expression Inducible expression Transduction Genetic modification 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Francisco Martín
    • 1
    Email author
  • María Tristán-Manzano
    • 1
  • Noelia Maldonado-Pérez
    • 1
  • Sabina Sánchez-Hernández
    • 1
  • Karim Benabdellah
    • 1
  • Marién Cobo
    • 1
  1. 1.Centre for Genomics and Oncological Research (GENYO)Pfizer/University of Granada/Andalusian Regional Government, PTS GranadaGranadaSpain

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