Department of Medicine, Renaissance School of Medicine at Stony Brook University.
Department of Medicine, Renaissance School of Medicine at Stony Brook University; Department of Physiology and Biophysics, Renaissance School of Medicine at Stony Brook University.
J Vis Exp. 2022 Jul 27(185). doi: 10.3791/64028.
The intestinal epithelium consists of a single layer of cells yet contains multiple types of terminally differentiated cells, which are generated by the active proliferation of intestinal stem cells located at the bottom of intestinal crypts. However, during events of acute intestinal injury, these active intestinal stem cells undergo cell death. Gamma irradiation is a widely used colorectal cancer treatment, which, while therapeutically efficacious, has the side effect of depleting the active stem cell pool. Indeed, patients frequently experience gastrointestinal radiation syndrome while undergoing radiotherapy, in part due to active stem cell depletion. The loss of active intestinal stem cells in intestinal crypts activates a pool of typically quiescent reserve intestinal stem cells and induces dedifferentiation of secretory and enterocyte precursor cells. If not for these cells, the intestinal epithelium would lack the ability to recover from radiotherapy and other such major tissue insults. New advances in lineage-tracing technologies allow tracking of the activation, differentiation, and migration of cells during regeneration and have been successfully employed for studying this in the gut. This study aims to depict a method for the analysis of cells within the mouse intestinal epithelium following radiation injury.
肠上皮由单层细胞组成,但包含多种终末分化细胞,这些细胞由位于肠隐窝底部的活跃增殖的肠干细胞产生。然而,在急性肠损伤事件中,这些活跃的肠干细胞会发生细胞死亡。伽马射线辐射是一种广泛应用于结直肠癌的治疗方法,虽然具有治疗效果,但也有耗尽活跃干细胞池的副作用。事实上,患者在接受放疗时常会出现胃肠道辐射综合征,部分原因是活跃的干细胞耗竭。肠隐窝中活跃肠干细胞的丧失会激活通常处于静止状态的储备肠干细胞池,并诱导分泌细胞和肠细胞前体细胞去分化。如果没有这些细胞,肠上皮将无法从放疗和其他重大组织损伤中恢复。谱系追踪技术的新进展允许在再生过程中追踪细胞的激活、分化和迁移,并已成功用于研究肠道中的这一过程。本研究旨在描述一种分析小鼠肠上皮细胞在辐射损伤后的方法。