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Isolation of Human Peripheral Blood T-Lymphocytes

  • Atish Kizhakeyil
  • Seow Theng Ong
  • Mobashar Hussain Urf Turabe Fazil
  • Madhavi Latha Somaraju Chalasani
  • Praseetha Prasannan
  • Navin Kumar Verma
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1930)

Abstract

Peripheral blood is the most common source of T-lymphocytes for in vitro culture. Here, we present a simple and standardized method for small- or large-scale isolation of viable T-lymphocytes and other mononuclear cells from fresh peripheral blood or buffy coat blood samples using the density gradient centrifugation. T-cells obtained using the protocol described here can be used for a variety of downstream analysis, including cellular, molecular, and functional assays.

Key words

T-Lymphocytes PBMC Density gradient Ficoll 

Notes

Acknowledgments

This work was supported by the Lee Kong Chian School of Medicine, Nanyang Technological University Singapore Start-Up Grant to N.K.V. and the Singapore Ministry of Education under its Singapore Ministry of Education (MOE) Academic Research Fund (AcRF) Tier 1 (2014-T1-001-141) and MOE-AcRF Tier 2 (MOE2017-T2-2-004). A.K. acknowledges Ph.D. fellowship provided by Lee Kong Chian School of Medicine, Nanyang Technological University Singapore.

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Copyright information

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

Authors and Affiliations

  • Atish Kizhakeyil
    • 1
  • Seow Theng Ong
    • 1
  • Mobashar Hussain Urf Turabe Fazil
    • 1
  • Madhavi Latha Somaraju Chalasani
    • 2
    • 1
  • Praseetha Prasannan
    • 1
  • Navin Kumar Verma
    • 3
  1. 1.Lymphocyte Signalling Research Laboratory, Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore
  2. 2.Autoimmunity and Inflammation ProgramHospital for Special SurgeryNew YorkUSA
  3. 3.Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore

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