Impact of preanalytical handling and timing for peripheral blood mononuclear cells isolation and rna studies: The experience of the interinstitutional multidisciplinary Biobank (BioBIM)

Raffaele Palmirotta, Maria Laura De Marchis, Giorgia Ludovici, Barbara Leone, Annalisa Savonarola, Cristiano Ialongo, Antonella Spila, Francesco De Angelis, Patrizia Ferroni, David Della Morte, Fiorella Guadagni

Research output: Contribution to journalArticle

13 Citations (Scopus)

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 PBMCs isolation prior to RNA extraction may have a significant impact on downstream molecular biological applications.

Original languageEnglish
Pages (from-to)90-98
Number of pages9
JournalInternational Journal of Biological Markers
Volume27
Issue number2
DOIs
StatePublished - Apr 1 2012

Fingerprint

Cell Separation
Blood Cells
Blood
RNA
Time delay
Blood Preservation
Agar Gel Electrophoresis
Electrophoresis
Sepharose
Nucleic Acids
Multicenter Studies
Real-Time Polymerase Chain Reaction
Gels
Temperature

Keywords

  • Biobank
  • RNA extraction
  • RNA quality
  • RNA quantity
  • Standard operating procedures

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cancer Research
  • Oncology
  • Pathology and Forensic Medicine

Cite this

Impact of preanalytical handling and timing for peripheral blood mononuclear cells isolation and rna studies : The experience of the interinstitutional multidisciplinary Biobank (BioBIM). / Palmirotta, Raffaele; De Marchis, Maria Laura; Ludovici, Giorgia; Leone, Barbara; Savonarola, Annalisa; Ialongo, Cristiano; Spila, Antonella; De Angelis, Francesco; Ferroni, Patrizia; Della Morte, David; Guadagni, Fiorella.

In: International Journal of Biological Markers, Vol. 27, No. 2, 01.04.2012, p. 90-98.

Research output: Contribution to journalArticle

Palmirotta, Raffaele ; De Marchis, Maria Laura ; Ludovici, Giorgia ; Leone, Barbara ; Savonarola, Annalisa ; Ialongo, Cristiano ; Spila, Antonella ; De Angelis, Francesco ; Ferroni, Patrizia ; Della Morte, David ; Guadagni, Fiorella. / Impact of preanalytical handling and timing for peripheral blood mononuclear cells isolation and rna studies : The experience of the interinstitutional multidisciplinary Biobank (BioBIM). In: International Journal of Biological Markers. 2012 ; Vol. 27, No. 2. pp. 90-98.
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