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

February 01, 2019 By:
  • Kofanova O
  • Bellora C
  • Aquilar-Quesada R
  • Bulla A
  • Panadero-Fajardo S
  • Keipes M
  • Shea K
  • Stone M
  • Lescuyer P
  • Betsou F.

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 degrees 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: CONCLUSION: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.

2019 Feb. J Immunol Methods.465:13-19. Epub 2018 Nov 26.
Other information