Impact of preanalytical handling and timing for peripheral blood mononuclear cells isolation and RNA studies: the experience of the Interinstitutional Multidisciplinary BioBank (BioBIM)

The International Journal of Biological Markers
Raffaele PalmirottaFiorella Guadagni

Abstract

Multicenter studies and biobanking projects require blood transportation from the participating center to a central collection or diagnostic laboratory. The impact of time delays between venous blood collection and peripheral blood mononuclear cells (PBMC) isolation prior to RNA extraction may affect the quality and quantity of isolated nucleic acids for genomic applications. Thus, standard operating procedure (SOP) optimization for the treatment of biological samples before RNA extraction is crucial in a biological repository. In order to define SOPs for whole blood preservation prior to RNA extraction, we sought to determine whether different blood storage times (0, 3, 6, 10, 24, and 30 hours) prior to PBMCs isolation and storage at -80°C, could affect the quality and quantity of extracted RNA. After spectrophotometric quantification, the quality and integrity of RNA were assessed by agarose gel electrophoresis, RNA integrity number and real time-PCR (RT-PCR).
Across the different time points we did not observe significant differences within the first 24 hours of blood storage at room temperature, while a significant loss in RNA yield and integrity was detected between 24 and 30 hours. We conclude that time delays before PBMC...Continue Reading

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Citations

May 17, 2015·Clinical Chemistry·Christina Ellervik, Jim Vaught
Jul 8, 2020·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Dahlia H TesfamichaelJessica C Pritchard
Jan 29, 2013·Neurological Sciences : Official Journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology·Raffaele PalmirottaFiorella Guadagni
Jul 23, 2013·The International Journal of Biological Markers·Raffaele PalmirottaFiorella Guadagni
Apr 2, 2015·Biopreservation and Biobanking·Gael HamotFay Betsou
Oct 4, 2012·The International Journal of Biological Markers·Umberto NanniFiorella Guadagni
Jun 2, 2015·Archives of Pathology & Laboratory Medicine·Jane H ZhouJeffrey N Myers

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