Precision-cut intestinal slices (PCIS) are used to study intestinal (patho)physiology, drug efficacy, toxicity, transport and metabolism ex vivo. One of the factors that limit the use of PCIS is a relatively short life-span. Moreover, culture-induced changes in cellular composition of PCIS remain largely uncharacterized. In this study, we demonstrated the epithelial cell heterogeneity in mouse and rat PCIS and its alterations during culture. In addition, we evaluated whether the presence of niche growth factors impacts the survival of PCIS epithelial cells. We showed that freshly prepared PCIS retained the main epithelial cell types, namely absorptive enterocytes, goblet cells, enteroendocrine cells, stem cells, transit-amplifying cells and Paneth cells. Once placed in culture, PCIS displayed progressive epithelial damage, and loss of these epithelial cell types. Cells comprising the intestinal stem cell niche were especially sensitive to the damage, and the addition of niche growth factors beneficially affected the survival of stem cells and transit-amplifying cells in PCIS during culture. In conclusion, this study provides new insights into the dynamic changes in cellular composition of epithelium in cultured PCIS, paving the way to future toxicological and pharmacological studies in an informed and reliable ex vivo setting.

精确切割的肠切片 (PCIS) 用于研究肠道（病理）生理学、药物功效、毒性、转运和体外代谢. 限制使用 PCIS 的因素之一是相对较短的使用寿命。此外，培养诱导的 PCIS 细胞组成的变化在很大程度上仍未得到表征。在这项研究中，我们证明了小鼠和大鼠 PCIS 的上皮细胞异质性及其在培养过程中的变化。此外，我们评估了生态位生长因子的存在是否会影响 PCIS 上皮细胞的存活。我们发现新鲜制备的 PCIS 保留了主要的上皮细胞类型，即吸收性肠细胞、杯状细胞、肠内分泌细胞、干细胞、转运放大细胞和 Paneth 细胞。一旦置于培养物中，PCIS 显示出进行性上皮损伤和这些上皮细胞类型的丢失。组成肠道干细胞生态位的细胞对损伤特别敏感，并且添加生态位生长因子有益地影响了培养过程中 PCIS 中干细胞和转运扩增细胞的存活。总之，这项研究为培养的 PCIS 中上皮细胞组成的动态变化提供了新的见解，为未来以知情和可靠的方式进行毒理学和药理学研究铺平了道路。离体环境。