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Intestinal stem cells remain viable after prolonged tissue storage

机译：肠道干细胞在长时间组织储存后仍然可行

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摘要

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.
机译：肠干细胞（ISCs）负责在正常稳态和损伤后的上皮再生。因此，它们具有显着的治疗潜力。但是，ISC是否可以存活组织储存是未知的。我们假设，尽管大多数上皮细胞可能死亡，但ISC将在4°C下至少24小时保持活力。为了探讨该假设，除去C57BL6 / J或LGR5-LacZ小鼠的Jejuna，并立即加工或在4℃下置于磷酸盐缓冲的盐水中。在24,30或48小时后进行上皮的延迟分离。在光学显微镜水平下，尽管对绒毛上皮细胞进行了广泛的凋亡，但小肠隐窝仍然是形态完整的30小时，并且通过LGR5-LacZ阳性识别ISC。电子显微镜表明，ISCS保留了24小时的高完整性。当通过流式细胞术评估时，ISC比其余的上皮细胞的变性更抗变性，包括相邻的群细胞，并且在所有时间点的可行性较高。培养的孤立的隐窝显示出在24小时后24小时后形成复合物的能力损失，7天培养后效率均超过80％。储存30小时，效率下降但保留了崭露头角的能力。我们得出结论，随着隔离延迟，ISC仍然可行并保留其增殖能力。相反，上皮的其余部分，包括Paneth细胞，表现出变性和编程的细胞死亡。如果这些发现在人体组织中储存，则在4°C的储存可能提供有价值的时间窗口，用于收集Crypts或ISCs进行治疗应用。

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  • 来源
    《Cell and Tissue Research》 |2013年第2期|共10页
  • 作者

    FullerM.K.; FaulkD.M.; SundaramN.; MaheM.M.; StoutK.M.; VonFurstenbergR.J.; SmithB.J.; McNaughtonK.K.; ShroyerN.F.; HelmrathM.A.; HenningS.J.;

  • 作者单位

    Department of Surgery University of North Carolina at Chapel Hill Chapel Hill NC United States;

    Department of Surgery Cincinnati Children's Hospital Medical Center Cincinnati OH United States;

    Department of Surgery Cincinnati Children's Hospital Medical Center Cincinnati OH United States;

    Department of Surgery Cincinnati Children's Hospital Medical Center Cincinnati OH United States;

    Department of Medicine and Cell Biology and Physiology University of North Carolina at Chapel Hill;

    Department of Medicine and Cell Biology and Physiology University of North Carolina at Chapel Hill;

    Department of Medicine and Cell Biology and Physiology University of North Carolina at Chapel Hill;

    Department of Medicine and Cell Biology and Physiology University of North Carolina at Chapel Hill;

    Department of Pediatrics Medical Center Cincinnati Children's Hospital Cincinnati OH United;

    Department of Surgery Cincinnati Children's Hospital Medical Center Cincinnati OH United States;

    University of North Carolina at Chapel Hill 4341 Medical Biomolecular Research Building (MBRB) CB;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;
  • 关键词

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

    机译：enceid培养;肠道细胞;光和电子显微镜;鼠标;耐贮藏;可行性;

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