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

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.