In Vitro Differentiation of CD4+ T Cell Effector and Regulatory Subsets

  • Jaclyn R. Espinosa
  • Joshua D. Wheaton
  • Maria CiofaniEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2111)


In vitro differentiation of naïve CD4+ T cells into effector and regulatory subsets offers a means to acquire large numbers of relatively homogeneous cell populations for experimentation. However, culture systems for T cell differentiation described in the literature vary widely in efficiency and complexity, limiting their comparison across studies. Here, we present a standardized and robust method for the isolation and in vitro differentiation of six CD4+ T cell subsets from mouse naïve T cells.

Key words

CD4 T cells In vitro differentiation Polarization Helper T cells Cytokines Th1 Th2 Th17 iTreg 



This work was supported by NIH grant R01 GM115474 to M.C.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Jaclyn R. Espinosa
    • 1
  • Joshua D. Wheaton
    • 1
  • Maria Ciofani
    • 1
    Email author
  1. 1.Department of ImmunologyDuke University Medical CenterDurhamUSA

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