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Itoh, Hiroshi; Yagi, Minoru; Hasebe, Ken; Fushida, Sachio; Tani, Takashi; Hashimoto, Takeji; Shimizu, Kouichi; Miwa, Koichi

doi:10.1023/a:1021049004188

Cited by 20 publications

(10 citation statements)

References 11 publications

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“…Moreover healthy ISCs at the crypt bases were observed both at the light microscopic (by Lgr5-LacZ ) and the EM level. Similar morphology of apoptotic villus loss with crypt preservation has also been noted in models of intestinal ischemia and reperfusion (Itoh, et al, 2002, Park and Haglund, 1992, Udassin, et al, 1994). Although no studies have looked specifically at the ISCs following ischemia and reperfusion, thymidine incorporation was noted at the base of the crypt in one study at the time of regeneration (Park and Haglund, 1992).…”

Section: Discussionsupporting

confidence: 72%

Intestinal stem cells remain viable after prolonged tissue storage

et al. 2013

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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, it is unknown whether ISCs can survive tissue storage. We hypothesized that, although the majority of epithelial cells may die, ISCs would remain viable for at least 24 h at 4°C. To explore this hypothesis, jejuni of C57Bl6/J or Lgr5-LacZ mice were removed and either processed immediately or placed in phosphate buffered saline (PBS) at 4°C. Delayed isolations of epithelia were 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 through 30 h and ISCs were identifiable via Lgr5-LacZ positivity. Electron microscopy showed that ISCs retain high integrity through 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. Culture of 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 with human tissue, storage at 4°C may offer a valuable temporal window for harvest of crypts or ISCs for therapeutic application.

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Section: Discussionsupporting

confidence: 72%

Intestinal stem cells remain viable after prolonged tissue storage

et al. 2013

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“…Mucosal damage is followed by transient rapid proliferation of intestinal crypt epithelial cells, migration of new cells from the crypt to the villus, and subsequent differentiation of villous cells to rebuild the proper structure of the epithelium [26], [27]. Studies have shown that the proliferation of intestinal epithelial cells occurs in the early phase after I/R, and immediate early genes, such as c -fos and c -jun, are involved in the proliferation of intestinal epithelial cells [28], [29]. Some other factors, such as histamine, histidine decarboxylase, and ornithine decarboxylase, were also reported to be involved in the regeneration of intestinal mucosa after I/R [30], [31].…”

Section: Discussionmentioning

confidence: 99%

The Jagged-2/Notch-1/Hes-1 Pathway Is Involved in Intestinal Epithelium Regeneration after Intestinal Ischemia-Reperfusion Injury

Qiu

Sun

et al. 2013

PLoS ONE

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BackgroundNotch signaling plays a critical role in the maintenance of intestinal crypt epithelial cell proliferation. The aim of this study was to investigate the role of Notch signaling in the proliferation and regeneration of intestinal epithelium after intestinal ischemia reperfusion (I/R) injury.MethodsMale Sprague-Dawley rats were subjected to sham operation or I/R by occlusion of the superior mesenteric artery (SMA) for 20 min. Intestinal tissue samples were collected at 0, 1, 2, 4, and 6 h after reperfusion. Proliferation of the intestinal epithelium was evaluated by immunohistochemical staining of proliferating nuclear antigen (PCNA). The mRNA and protein expression levels of Notch signaling components were examined using Real-time PCR and Western blot analyses. Immunofluorescence was also performed to detect the expression and location of Jagged-2, cleaved Notch-1, and Hes-1 in the intestine. Finally, the γ-secretase inhibitor DAPT and the siRNA for Jagged-2 and Hes-1 were applied to investigate the functional role of Notch signaling in the proliferation of intestinal epithelial cells in an in vitro IEC-6 culture system.ResultsI/R injury caused increased intestinal crypt epithelial cell proliferation and increased mRNA and protein expression of Jagged-2, Notch-1, and Hes-1. The immunofluorescence results further confirmed increased protein expression of Jagged-2, cleaved Notch-1, and Hes-1 in the intestinal crypts. The inhibition of Notch signaling with DAPT and the suppression of Jagged-2 and Hes-1 expression using siRNA both significantly inhibited the proliferation of IEC-6 cells.ConclusionThe Jagged-2/Notch-1/Hes-1 signaling pathway is involved in intestinal epithelium regeneration early after I/R injury by increasing crypt epithelial cell proliferation.

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“…Studies have shown that intestinal epithelial cell apoptosis is a significant contributor to mucosal barrier dysfunction. Immediate early genes, such as c- fos and c -jun are involved in the apoptosis of intestinal epithelial cells [22,23]. Some other factors, such as p38 MAPK, IL-6, HGF, and KGF, were also reported to be involved in the apoptosis of intestinal epithelial cells after I/R injury [24–27].…”

Section: Discussionmentioning

confidence: 99%

“…Inhibition of Notch signaling activation by γ-secretase inhibitors also lead to disturbed intestinal homeostasis [28–30]. The published studies have shown that the PCNA-positive cells increased at 1 to 6 h after reperfusion and TUNEL-positive cells increased later than PCNA-positive cells in a rat I/R model [22,23,31]. Our previous studies have investigated the role of Notch signaling in the proliferation of intestinal epithelial cells [19,32].…”

Section: Discussionmentioning

confidence: 99%

The Canonical Notch Signaling Was Involved in the Regulation of Intestinal Epithelial Cells Apoptosis after Intestinal Ischemia/Reperfusion Injury

Zhang¹,

Cheng²,

Xiao³

et al. 2014

IJMS

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Notch signaling plays a critical role in the maintenance of intestinal homeostasis. The aim of the present study was to investigate the role of Notch signaling in the apoptosis of intestinal epithelial cells after intestinal ischemia reperfusion (I/R) injury. Male C57BL/6 mice were subjected to sham operation or I/R injury. Intestinal tissue samples were collected at 12 h after reperfusion. TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling) staining showed that intestinal I/R injury induced significantly increased apoptosis of intestinal epithelial cells. Meanwhile, the mRNA expression of Jagged1, DLL1, Notch2, and Hes5, and protein expression of NICD2 and Hes5 were increased significantly after I/R injury in intestinal epithelial cells. In an in vitro IEC-6 culture model, flow cytometry analyses showed that inhibition of Notch signaling by γ-secretase inhibitor DAPT and the suppression of Hes5 expression using siRNA both significantly increased the apoptosis of IEC-6 cells under the condition of hypoxia/reoxygenation (H/R). In conclusion, the Notch2/Hes5 signaling pathway was activated and involved in the regulation of intestinal epithelial cells apoptosis in intestinal I/R injury.

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Untitled

Cited by 20 publications

References 11 publications

Intestinal stem cells remain viable after prolonged tissue storage

Intestinal stem cells remain viable after prolonged tissue storage

The Jagged-2/Notch-1/Hes-1 Pathway Is Involved in Intestinal Epithelium Regeneration after Intestinal Ischemia-Reperfusion Injury

The Canonical Notch Signaling Was Involved in the Regulation of Intestinal Epithelial Cells Apoptosis after Intestinal Ischemia/Reperfusion Injury

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