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Effect of delayed cell processing and cryopreservation on immunophenotyping in multicenter population studies

Bharat Thyagarajan1, Helene Barcelo1, Eileen Crimmins2

  • 1Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN, United States.

Journal of Immunological Methods
|September 18, 2018
PubMed

Insights

Delayed cell processing and cryopreservation impact immune cell percentages in large studies. Blood processing within 48 hours is feasible for immunophenotyping, though some cell subsets are sensitive to combined delays and freezing.

Area of Science:

  • Immunology
  • Flow Cytometry
  • Biotechnology

Background:

  • Delayed cell processing and cryopreservation introduce variability in immunophenotyping.
  • Accurate immune cell subset quantification is crucial for large population studies.

Purpose of the Study:

  • To assess the impact of delayed processing (up to 72 hours) and cryopreservation on immunophenotyping results.
  • To evaluate the combined effects of delayed processing and cryopreservation on various immune cell subsets.

Main Methods:

  • Pilot study involving 20 volunteers.
  • Blood samples processed immediately, after delays (up to 72h), and after cryopreservation.
  • Immunophenotyping of 31 cell subsets (T cells, B cells, NK cells, monocytes, dendritic cells) using EDTA and CPT tubes.

Main Results:

  • Delayed processing or cryopreservation alone increased B and NK cell percentages.
  • Combined delayed processing and cryopreservation increased T cell percentages but decreased NK cell percentages.
  • B cell increases were significant with 24-48h delays; monocytes and dendritic cells remained unaffected.

Conclusions:

  • Immunophenotyping for large population studies is feasible within 48 hours of collection.
  • Certain immune cell subsets show differential sensitivity to combined delayed processing and cryopreservation.
  • Standardized protocols are essential to mitigate variability in large-scale immunophenotyping studies.

Keywords:
Cell processingCohort studyCryopreservationImmunophenotyping

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