IL8 and EDEM3 gene expression ratio indicates peripheral blood mononuclear cell (PBMC) quality

Area of Science:

• Biomedical Science
• Molecular Biology
• Clinical Diagnostics

Background:

• Preanalytical variables significantly affect the accuracy and reproducibility of results derived from peripheral blood mononuclear cells (PBMC).
• Maintaining PBMC quality is crucial for reliable downstream analytical outcomes.
• Standardization of PBMC isolation and handling is essential for consistent research and clinical applications.

Purpose of the Study:

• To develop and validate a metric for assessing PBMC quality.
• To evaluate the impact of pre-centrifugation delays on PBMC gene expression.
• To establish a method for objective PBMC sample qualification.

Main Methods:

• Peripheral blood mononuclear cells (PBMC) were isolated from EDTA and citrate-anticoagulated blood samples.
• Gene expression changes in PBMC-derived RNA were analyzed following extended pre-centrifugation delays at 4°C and room temperature (RT).
• Taqman RT-qPCR was employed to assess the diagnostic performance of two target genes in identifying samples with extended pre-centrifugation times.

Main Results:

• A novel PBMC preanalytical score was established as a gene expression metric.
• This score assesses PBMC quality in relation to pre-centrifugation delay and anticoagulant type.
• The PBMC preanalytical score effectively identifies sample quality variations.

Conclusions:

• The developed PBMC preanalytical score offers an objective method for qualifying PBMC samples.
• This metric is valuable for downstream applications sensitive to blood pre-centrifugation delays.
• Implementation of the PBMC preanalytical score can enhance the reliability of PBMC-based analyses.