title: Applicability of Under Vacuum Fresh Tissue Sealing and Cooling to Omics Analysis of Tumor Tissues.
abstract: BACKGROUND
  Biobanks of frozen human normal and malignant tissues represent a valuable source for "omics" analysis in translational cancer research and molecular pathology. However, the success of molecular and cellular analysis strongly relies on the collection, handling, storage procedures, and quality control of fresh human tissue samples.
  
  
  OBJECTIVE
  We tested whether under vacuum storage (UVS) effectively preserves tissues during the time between surgery and storage for "omics" analyses.
  
  
  METHODS
  Normal and matched tumor specimens, obtained from 16 breast, colon, or lung cancer patients and 5 independent mesenchymal tumors, were dissected within 20 minutes from surgical excision and divided in three to five aliquots; for each tissue sample, one aliquot was snap-frozen in liquid nitrogen (defined as baseline or T0 samples), and the other portions were sealed into plastic bags and kept at 4°C for 1, 24, 48, or 72 hours under vacuum and then frozen. The tissue and molecular preservation under vacuum was evaluated over time in terms of histomorphology, transcription (Illumina microarrays), protein (surface-enhanced laser desorption/ionization-time of flight/mass spectrometry and Western blot), and metabolic profile (nuclear magnetic resonance spectroscopy).
  
  
  RESULTS
  Tissue morphology, Mib-1, and vimentin immunostaining were preserved over time without signs of tissue degradation. Principal variance component analysis showed that time of storage had a minimal effect on gene expression or the proteome, but affected the preservation of some metabolites to a greater extent. UVS did not impact the RNA and protein integrity or specific phosphorylation sites on mTOR and STAT3. Measurement of metabolites revealed pronounced changes after 1 hour of storage.
  
  
  CONCLUSIONS
  Our results show that UVS can preserve tissue specimens for histological, transcriptomic, and proteomic examinations up to 48 hours and possibly longer, whereas it has limitations for metabolomic applications.
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Silvia
authors.initials: S
authors.lastname: Veneroni
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Matteo
authors.initials: M
authors.lastname: Dugo
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Daidone
authors.affiliation: 2 Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità , Rome, Italy .
authors.forename: Egidio
authors.initials: E
authors.lastname: Iorio
authors.affiliation: 3 Department of Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Barbara
authors.initials: B
authors.lastname: Valeri
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Patrizia
authors.initials: P
authors.lastname: Pinciroli
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Maida
authors.initials: M
authors.lastname: De Bortoli
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Edoardo
authors.initials: E
authors.lastname: Marchesi
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Patrizia
authors.initials: P
authors.lastname: Miodini
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Elena
authors.initials: E
authors.lastname: Taverna
authors.affiliation: 2 Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità , Rome, Italy .
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ricci
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Silvana
authors.initials: S
authors.lastname: Canevari
authors.affiliation: 3 Department of Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .|4 Department of Biomedical and Clinical Sciences "Luigi Sacco," Università degli Studi di Milano , Milan, Italy .
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Pelosi
authors.affiliation: 1 Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori , Milan, Italy .
authors.forename: Italia
authors.initials: I
authors.lastname: Bongarzone
mesh_terms: D005260:Female; D006801:Humans; D008297:Male; D055432:Metabolomics; D009369:Neoplasms / Q000473:pathology*; D040901:Proteomics; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods*; D014015:Tissue Banks / Q000592:standards*; D057170:Translational Research, Biomedical; D014618:Vacuum
keywords: biobanking; high-throughput analysis; histopathology; ischemic time; tissue preservation; under-vacuum