Ex vivo culture of intestinal crypt organoids as a model system for assessing cell death induction in intestinal epithelial cells and enteropathy.
Ex vivo culture of intestinal crypt organoids as a model system for assessing cell death induction in intestinal epithelial cells and enteropathy.
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DOI：
10.1038/cddis.2014.183
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发表时间：
2014-05-15
影响因子：
9
通讯作者：
中科院分区：
文献类型：
--
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肠上皮细胞（IEC）不仅在营养吸收方面具有关键功能，而且还是我们身体与外界之间的物理屏障。由于免疫系统接触肠道菌群的化合物，上皮细胞的损伤和死亡导致这种屏障功能的破坏和炎症。肠上皮损伤通常与各种炎症性疾病、化疗和放疗以及药物介导的毒性有关。直到最近，由于原代 IEC 的离体存活率很低，药物和治疗方法的肠上皮损伤活性只能在动物模型体内进行测试。肠隐窝干细胞的三维培养和类器官的生长为离体培养和研究 IEC 提供了新的可能性。在这里，我们证明肠道类器官是研究 IEC 细胞死亡和存活的有用且生理相关的模型系统。我们进一步描述了许多基于显微镜和比色的方法来监测和评分肠道类器官的存活和死亡。最后，通过比较从基因缺陷小鼠和野生型小鼠中分离出的类器官，可以研究特定基因在 IEC 死亡调节中的作用。由于其相似的结构和行为，肠类器官可以作为一种有趣且生理相关的替代系统，用于大中型体外测试肠上皮损伤药物和毒素，以及用于研究细胞死亡途径。
Intestinal epithelial cells (IECs) not only have a critical function in the absorption of nutrients, but also act as a physical barrier between our body and the outside world. Damage and death of the epithelial cells lead to the breakdown of this barrier function and inflammation due to access of the immune system to compounds of the intestinal flora. Intestinal epithelial damage is frequently associated with various inflammatory disorders, chemo- and radiotherapy as well as drug-mediated toxicity. Until recently, intestinal epithelial-damaging activities of drugs and treatments could be tested only in vivo in animal models because of the poor survival rate of primary IECs ex vivo. The three-dimensional culture and outgrowth of intestinal crypt stem cells into organoids have offered new possibilities to culture and study IECs ex vivo. Here we demonstrate that intestinal organoids are a useful and physiologically relevant model system to study cell death and survival in IECs. We further describe a number of microscopy-based as well as colorimetric methods to monitor and score survival and death of intestinal organoids. Finally, the comparison of organoids isolated from gene-deficient mice and wild-type mice allows investigating the role of specific genes in the regulation of IEC death. Owing to their comparable structure and behavior, intestinal organoids may serve as an interesting and physiologically relevant surrogate system for large- and mid-scale in vitro testing of intestinal epithelium-damaging drugs and toxins, and for the investigation of cell death pathways.
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DOI：
10.1073/pnas.1432897100
发表时间：
2003-07-08
影响因子：
11.1
作者：
Marini, M;Bamias, G;Cominelli, F
通讯作者：
Cominelli, F
影响因子：
6.1
作者：
Kalive, Madhavi;Zhang, Wen;Capco, David G.
通讯作者：
Capco, David G.
影响因子：
2.2
作者：
Sala, Frederic G.;Kunisaki, Shaun M.;Grikscheit, Tracy C.
通讯作者：
Grikscheit, Tracy C.
影响因子：
7.3
作者：
Lau KS;Juchheim AM;Cavaliere KR;Philips SR;Lauffenburger DA;Haigis KM
通讯作者：
Haigis KM
影响因子：
2.3
作者：
Bowen, Joanne M.;Gibson, Rachel J.;Keefe, Dorothy M. K.
通讯作者：
Keefe, Dorothy M. K.