Precision-cut rat, mouse, and human intestinal slices as novel models for the early-onset of intestinal fibrosis

Abstract

Intestinal fibrosis (IF) is a major complication of inflammatory bowel disease. IF research is limited by the lack of relevant in vitro and in vivo models. We evaluated precision-cut intestinal slices (PCIS) prepared from human, rat, and mouse intestine as ex vivo models mimicking the early-onset of (human) IF. Precision-cut intestinal slices prepared from human (h), rat (r), and mouse (m) jejunum, were incubated up to 72 h, the viability of PCIS was assessed by ATP content and morphology, and the gene expression of several fibrosis markers was determined. The viability of rPCIS decreased after 24 h of incubation, whereas mPCIS and hPCIS were viable up to 72 h of culturing. Furthermore, during this period, gene expression of heat shock protein 47 and plasminogen activator inhibitor 1 increased in all PCIS in addition to augmented expression of synaptophysin in hPCIS, fibronectin (Fn2) and TGF-β1 in rPCIS, and Fn2 and connective tissue growth factor (Ctgf) in mPCIS. Addition of TGF-β1 to rPCIS or mPCIS induced the gene expression of the fibrosis markers Pro-collagen1a1, Fn2, and Ctgf in both species. However, none of the fibrosis markers was further elevated in hPCIS. We successfully developed a novel ex vivo model that can mimic the early-onset of fibrosis in the intestine using human, rat, and mouse PCIS. Furthermore, in rat and mouse PCIS, TGF-β1 was able to even further increase the gene expression of fibrosis markers. This indicates that PCIS can be used as a model for the early-onset of IF.

Transcriptomic characterization of culture-associated changes in murine and human precision-cut tissue slices

“…Recently, the application of PCTS was further extended to study the mechanism of fibrosis, which is characterized by the excess deposition of extracellular matrix. It has been shown that PCTS from different organs develop inflammatory and fibrogenic responses during culture, making PCTS a suitable model for fibrosis and analysing the effect of antifibrotic compounds (Kasper et al 2004;Westra et al 2014Westra et al , 2016Pham et al 2015;Stribos et al 2016;Alsafadi et al 2017;Luangmonkong et al 2017Luangmonkong et al , 2018. Furthermore, PCTS generated from human diseased organs allow the investigation of fibrosis in situ, as we previously showed for human cirrhotic liver PCTS (Luangmonkong et al 2017).…”

Precision-Cut Kidney Slices as a Tool to Understand the Dynamics of Extracellular Matrix Remodeling in Renal Fibrosis

“…8 However, most of these models use healthy tissues from rats or mice and stimulate them with TGF-β1 to induce a fibrogenic response. [15][16][17] In a recent study, the use of PCTS from human kidneys has been proposed to study the mechanisms of fibrosis. 18 However also in this experimental approach, fibrosis is induced in already cultured slices by means of TGF-β stimulation.…”

Infection of porcine precision cut intestinal slices by transmissible gastroenteritis coronavirus demonstrates the importance of the spike protein for enterotropism of different virus strains

“…These slices are produced to stay vital in culture and have been already used to study the intestinal tissue in smaller species like rodents (Niu et al, 2014). So far they have been established in rat, mouse, and human jejunum (Pham et al, 2015). Here for the first time, we established porcine jejunum PCIS and demonstrate that they can be infected by different TGEV strains.…”