Applicability of under Vacuum Fresh Tissue Sealing and Cooling to Omics Analysis of Tumor Tissues

Silvia Veneroni, Matteo Dugo, Maria Grazia Daidone, Egidio Iorio, Barbara Valeri, Patrizia Pinciroli, Maida De Bortoli, Edoardo Marchesi, Patrizia Miodini, Elena Taverna, Alessandro Ricci, Silvana Canevari, Giuseppe Pelosi, Italia Bongarzone

Research output: Contribution to journalArticlepeer-review

Abstract

Context: 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. Design: 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.

Original languageEnglish
Pages (from-to)480-490
Number of pages11
JournalBiopreservation and Biobanking
Volume14
Issue number6
DOIs
Publication statusPublished - Dec 1 2016

Keywords

  • Biobanking
  • high-throughput analysis
  • histopathology
  • ischemic time
  • tissue preservation
  • under-vacuum

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Cell Biology

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