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

Olga Kofanova, Camille Bellora, Rocio Aguilar Quesada, Alexandre Bulla, Sonia Panadero-Fajardo, Marc Keipes, Kathi Shea, Mars Stone, Pierre Lescuyer, Fay Betsou*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)

Abstract

Background: Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs). Methods: PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. We used Taqman RTqPCR to evaluate the combination of two target genes for their “diagnostic performance” in identifying EDTA and citrate-anticoagulated PBMC samples with extended pre-centrifugation times. Results: We established the PBMC preanalytical score, a gene expression metric to asses the PBMC quality related to the pre-centrifugation delay at room temperature for different anticoagulants. The PBMC preanalytical score measurement can identify: (1) EDTA PBMC samples or RNA extracted from these PBMCs with RT precentrifugation times >48 h with 98% sensitivity and 87% specificity at a cutoff of 57. (2) citrate PBMC samples or RNA extracted from these PBMCs with RT precentrifugation times of >48 h with 92% sensitivity and 84% specificity at a cutoff of 348. Conclusion: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.

Original languageEnglish
Pages (from-to)13-19
Number of pages7
JournalJournal of Immunological Methods
Volume465
DOIs
Publication statusPublished - Feb 2019

Keywords

  • PBMC
  • Preanalytical
  • Quality control

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