Asal Melis, Thon Maria, Waaijman Taco, Bontkes Hetty J, van Vliet Sandra J, Mebius Reina E, Gibbs Susan
Department of Molecular Cell Biology and Immunology, Amsterdam UMC Location Vrije Universiteit Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
Amsterdam Institute for Immunology and Infectious Diseases, 1081 HZ Amsterdam, The Netherlands.
Biomedicines. 2024 Dec 21;12(12):2913. doi: 10.3390/biomedicines12122913.
: The barrier properties of the human small intestine play a crucial role in regulating digestion, nutrient absorption and drug metabolism. Current in vitro organotypic models consist only of an epithelium, which does not take into account the possible role of stromal cells such as fibroblasts or the extracellular matrix (ECM) which could contribute to epithelial barrier properties. Therefore, the aim of this study was to determine whether these stromal cells or ECM were beneficial or detrimental to barrier function when incorporated into an organotypic human small intestine model. : Intestinal epithelial cell lines or primary cell organoids derived from the epithelial stem cells of the small intestine were cultivated either on a porous Transwell membrane (epithelial model) or on a primary small intestinal stromal cell-populated collagen-fibrin hydrogel (full thickness model). : Both models expressed villin (enterocytes), lysozyme (Paneth cells), Ki67 (proliferative cells) and zonula occludens-1 (tight junctions). The polarized epithelial barriers of the full thickness models demonstrated a significant decrease in transepithelial electrical resistance (TEER) with values comparable to that found in the native small intestine in contrast to the higher TEER values observed in the epithelial models. This correlated to an increase in secreted zonulin, a regulator of intestine permeability, in the full thickness models. The decreased TEER values were due to both the stromal cells and the choice of the hydrogel versus the Transwell membrane. Moreover, erythropoietin and epithelial growth factor secretion, which have roles in regulating barrier integrity, directly correlated with the changes in TEER and permeability. : This study emphasizes the importance of different cell types being incorporated into small intestine models and, also, the influence of the scaffold or matrix used.
人类小肠的屏障特性在调节消化、营养吸收和药物代谢方面起着至关重要的作用。目前的体外器官型模型仅由上皮细胞组成，没有考虑到成纤维细胞等基质细胞或细胞外基质（ECM）可能对上皮屏障特性产生的作用。因此，本研究的目的是确定将这些基质细胞或细胞外基质纳入器官型人类小肠模型时，它们对屏障功能是有益还是有害。：从小肠上皮干细胞衍生的肠上皮细胞系或原代细胞类器官，分别培养在多孔Transwell膜上（上皮模型）或原代小肠基质细胞填充的胶原-纤维蛋白水凝胶上（全层模型）。：两种模型均表达绒毛蛋白（肠细胞）、溶菌酶（潘氏细胞）、Ki67（增殖细胞）和闭合蛋白-1（紧密连接）。与上皮模型中观察到的较高跨上皮电阻（TEER）值相比，全层模型的极化上皮屏障的TEER值显著降低，其值与天然小肠中的值相当。这与全层模型中分泌的zonulin（一种肠道通透性调节剂）增加相关。TEER值降低是由于基质细胞以及水凝胶与Transwell膜的选择所致。此外，在调节屏障完整性方面起作用的促红细胞生成素和上皮生长因子的分泌，与TEER和通透性的变化直接相关。：本研究强调了将不同细胞类型纳入小肠模型的重要性，以及所用支架或基质的影响。
