Development of Feline Ileum- and Colon-Derived Organoids and Their Potential Use to Support Feline Coronavirus Infection

Abstract

Feline coronaviruses (FCoVs) infect both wild and domestic cat populations world-wide. FCoVs present as two main biotypes: the mild feline enteric coronavirus (FECV) and the fatal feline infectious peritonitis virus (FIPV). FIPV develops through mutations from FECV during a persistence infection. So far, the molecular mechanism of FECV-persistence and contributing factors for FIPV development may not be studied, since field FECV isolates do not grow in available cell culture models. In this work, we aimed at establishing feline ileum and colon organoids that allow the propagation of field FECVs. We have determined the best methods to isolate, culture and passage feline ileum and colon organoids. Importantly, we have demonstrated using GFP-expressing recombinant field FECV that colon organoids are able to support infection of FECV, which were unable to infect traditional feline cell culture models. These organoids in combination with recombinant FECVs can now open the door to unravel the molecular mechanisms by which FECV can persist in the gut for a longer period of time and how transition to FIPV is achieved.

Organoids: a promising new in vitro platform in livestock and veterinary research

“…This suggested that mesenchymal-like cell types are essential for cat enteroid growth. Recently, Tekes et al reported to have generated colon-derived organoids and studied host–pathogen interactions and immune response against feline enteric coronavirus [ 100 ].…”

“…The respiratory, gastrointestinal, urogenital, and mammary epithelia, along with the skin, are the most important sites of contact with the outside environment, with intimate contacts with the respective microbiomes of these organs, including potential pathogens. While interaction with microbes has mostly been studied in organoid models of intestinal epithelia of different species [ 55 , 56 , 59 , 61 – 64 , 71 – 73 , 83 , 84 , 91 , 100 , 103 ], similar studies could be done in organoid models as have been done in human and rodents for the lung [ 121 ] and the mammary gland [ 122 ]. In the gut, the epithelium is constantly screening the contents of the intestinal lumen [ 123 , 124 ].…”

“…In cattle, Ellis et al reported long-term culture of bovine 3D mammary gland organoids that recapitulated [145] Bile duct [146] Liver [147][148][149][150][151] Urinary bladder [152] Testis [44,45] Chicken Nervous and lymphoid tissue [47] Skeletal muscles [46] Cattle Arteries [153][154][155][156][157] Articular cartilage [158,159] Intestine [160] Mammary gland [48][49][50][51][52] Parathyroid gland [161][162][163][164][165][166] Dog Intestine [167] Prostate gland [98] Urinary bladder [99] Kidney [97] Intestine [55,91] Liver [96] Cat Intestine [55,100] Liver [101,102] Horse Intestine [55,103] glandular duct morphology and function [48]. Their culture methods included the use of ECM (collagen) and growth factors, such as IGF1, TGFβ, and mammary gland extract.…”

The Organoid Platform: Promises and Challenges as Tools in the Fight against COVID-19

“…ASC-derived intestinal organoids have been derived from non-human species, including bat and feline. This has allowed the propagation of strains of coronaviruses directly from bats ( Tekes et al., 2020 ; Zhou et al., 2020 ). These efforts highlight how organoids can be exploited to study (species-specific) viruses with zoonotic potential.…”

Enteropathogenic Infections: Organoids Go Bacterial

“…Considering the presumable zoonotic background of SARS-CoV-2, researchers have for the first time established intestinal organoids from Chinese horseshoe bats suspected to be one of the natural reservoirs [ 112 ]. With the intestinal organoid culturing protocol also being applicable to other non-human mammalian species such as the cow [ 113 ], pig [ 113 , 114 ], dog [ 115 ], and cat [ 116 ], important strides have been made to consolidate our current pathophysiological understanding of zoonotic diseases. Combining the findings derived from intestinal organoids spanning different species will certainly be of added value for characterizing a broad spectrum of common zoonotic bacterial pathogens affecting the intestinal tract.…”