Cell and Tissue Research

, Volume 354, Issue 2, pp 441–450

Intestinal stem cells remain viable after prolonged tissue storage

  • Megan K. Fuller
  • Denver M. Faulk
  • Nambirajan Sundaram
  • Maxime M. Mahe
  • Kara M. Stout
  • Richard J. von Furstenberg
  • Brian J. Smith
  • Kirk K. McNaughton
  • Noah F. Shroyer
  • Michael A. Helmrath
  • Susan J. Henning
Regular Article
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Abstract

Intestinal stem cells (ISCs) are responsible for renewal of the epithelium both during normal homeostasis and following injury. As such, they have significant therapeutic potential. However, whether ISCs can survive tissue storage is unknown. We hypothesize that, although the majority of epithelial cells might die, ISCs would remain viable for at least 24 h at 4 °C. To explore this hypothesis, jejuna of C57Bl6/J or Lgr5-LacZ mice were removed and either processed immediately or placed in phosphate-buffered saline at 4 °C. Delayed isolation of epithelium was performed after 24, 30, or 48 h storage. At the light microscope level, despite extensive apoptosis of villus epithelial cells, small intestinal crypts remained morphologically intact for 30 h and ISCs were identifiable via Lgr5-LacZ positivity. Electron microscopy showed that ISCs retained high integrity for 24 h. When assessed by flow cytometry, ISCs were more resistant to degeneration than the rest of the epithelium, including neighboring Paneth cells, with higher viability across all time points. Cultured isolated crypts showed no loss of capacity to form complex enteroids after 24 h tissue storage, with efficiencies after 7 days of culture remaining above 80 %. By 30 h storage, efficiencies declined but budding capability was retained. We conclude that, with delay in isolation, ISCs remain viable and retain their proliferative capacity. In contrast, the remainder of the epithelium, including the Paneth cells, exhibits degeneration and programmed cell death. If these findings are recapitulated in human tissue, storage at 4 °C might offer a valuable temporal window for the harvesting of crypts or ISCs for therapeutic application.

Keywords

Intestinal stem cells Light and electron microscopy Viability Resistance to storage Enteroid culture Mouse 

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Megan K. Fuller
    • 1
  • Denver M. Faulk
    • 3
  • Nambirajan Sundaram
    • 3
  • Maxime M. Mahe
    • 3
  • Kara M. Stout
    • 2
  • Richard J. von Furstenberg
    • 2
  • Brian J. Smith
    • 2
  • Kirk K. McNaughton
    • 2
  • Noah F. Shroyer
    • 4
  • Michael A. Helmrath
    • 3
  • Susan J. Henning
    • 5
  1. 1.Department of SurgeryUniversity of North Carolina at Chapel HillChapel HillUSA
  2. 2.Department of Medicine and Cell Biology & PhysiologyUniversity of North Carolina at Chapel HillChapel HillUSA
  3. 3.Department of SurgeryCincinnati Children’s Hospital Medical CenterCincinnatiUSA
  4. 4.Department of Pediatrics: Division of Gastroenterology, Hepatology & NutritionCincinnati Children’s Hospital Medical CenterCincinnatiUSA
  5. 5.University of North Carolina at Chapel HillChapel HillUSA

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