A Practical Cryopreservation and Staining Protocol for Immunophenotyping in Population Studies

Helene Barcelo, Jessica Faul, Eileen Crimmins, Bharat Thyagarajan

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21 Scopus citations


Large population-based cohort studies, through their prospective collection of a broad range of health information, represent an invaluable resource for novel insights into the pathogenesis of human diseases. Collection and cryopreservation of viable cells from blood samples is becoming increasingly common in large cohorts as these cells are a valuable resource for immunophenotyping and functional studies. The cryopreservation of peripheral blood mononuclear cells (PBMCs), thawing, and immunophenotyping protocols used to immunophenotype 9938 participants in the Health and Retirement Study (HRS) are described. The extensive quality control involved in a large-scale immunophenotyping epidemiological study is also outlined. The existing literature on the effect of cryopreservation on various immune cell subsets including T, B, NK cells, monocytes, and dendritic cells is provided.

Original languageEnglish (US)
Article numbere35
JournalCurrent protocols in cytometry
Issue number1
StatePublished - Apr 2018

Bibliographical note

Funding Information:
This work was funded by a grant from the National Institute of Aging (U01 AG009740). The authors would like to thank Sharon Minnerath, Vicky Makky, and Eileen Studt for assistance with cryopreservation and Zach Flaten, Devon Hunter‐Schlichting, Erin Cassidy, Paige Hartman, Tori Contreras, and Ramya Ramasubramian for assistance with immunophenotyping of cryopreserved samples.

Publisher Copyright:
Copyright © 2018 John Wiley & Sons, Inc.


  • cryopreservation
  • flow cytometry
  • immunophenotyping
  • peripheral blood mononuclear cells (PBMCs)
  • quality control
  • thawing


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