title: Effects of Transport and Storage Conditions on Gene Expression in Blood Samples.
abstract: BACKGROUND
  Inappropriate handling of blood samples might induce or repress gene expression and/or lead to RNA degradation affecting downstream analysis. In particular, sample transport is a critical step for biobanking or multicenter studies because of uncontrolled variables (i.e., unstable temperature). We report the results of a pilot study implemented within the EC funded SPIDIA project, aimed to investigate the role of transport and storage of blood samples containing and not containing an RNA stabilizer.
  
  
  METHODS
  Blood was collected from a single donor both in EDTA and in PAXgene Blood RNA tubes. Half of the samples were sent to a second laboratory both at room temperature and at 4°C, whereas the remaining samples were stored at room temperature and at 4°C. Gene expression of selected genes (c-FOS, IL-1β, IL-8, and GAPDH) known to be induced or repressed by ex vivo blood handling and of blood-mRNA quality biomarkers identified and validated within the SPIDIA project, which allow for monitoring changes in unstabilized blood samples after collection and during transport and storage, were analyzed by RT-qPCR.
  
  
  RESULTS
  If the shipment of blood in tubes not containing RNA stabilizer is not performed under a stable condition, gene profile studies can be affected by the effects of transport. Moreover, also controlled temperature shipment (4°C) can influence the expression of specific genes if blood is collected in tubes not containing a stabilizer.
  
  
  CONCLUSIONS
  The use of dedicated biomarkers or time course experiments should be performed in order to verify potential bias on gene expression analysis due to sample shipment and storage conditions. Alternatively, the use of RNA stabilizer containing tubes can represent a reliable option to avoid ex vivo RNA changes.
authors.affiliation: 1 Department of Biomedical, Experimental and Clinical Sciences, University of Florence , Italy .
authors.forename: Francesca
authors.initials: F
authors.lastname: Malentacchi
authors.affiliation: 2 Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Sara
authors.initials: S
authors.lastname: Pizzamiglio
authors.affiliation: 3 Qiagen GmbH , Hilden, Germany .
authors.forename: Ralf
authors.initials: R
authors.lastname: Wyrich
authors.affiliation: 2 Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Paolo
authors.initials: P
authors.lastname: Verderio
authors.affiliation: 2 Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Chiara
authors.initials: C
authors.lastname: Ciniselli
authors.affiliation: 1 Department of Biomedical, Experimental and Clinical Sciences, University of Florence , Italy .
authors.forename: Mario
authors.initials: M
authors.lastname: Pazzagli
authors.affiliation: 1 Department of Biomedical, Experimental and Clinical Sciences, University of Florence , Italy .
authors.forename: Stefania
authors.initials: S
authors.lastname: Gelmini
mesh_terms: D001769:Blood; D015870:Gene Expression; D006801:Humans; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature