Human ileal organoid model recapitulates clinical incidence of diarrhea associated with small molecule drugs

Yang

et al. 2021

Animals

Intestinal organoids offer great promise for disease-modelling-based host–pathogen interactions and nutritional research for feed efficiency measurement in livestock and regenerative medicine for therapeutic purposes. However, very limited studies are available on the functional characterisation and three-dimensional (3D) expansion of adult stem cells in livestock species compared to other species. Intestinal crypts derived from intestinal organoids under a 3D culture system from the small intestine in adult bovine were successfully established and characterised for functionality testing, including the cellular potentials and genetic properties based on immunohistochemistry, immunocytochemistry, epithelial barrier permeability assay, QuantSeq 3′ mRNA-Seq. data and quantitative reverse transcription-polymerase chain reaction. Intestinal organoids were long-term cultivated over several passages of culture without loss of the recapitulating capacity of crypts, and they had the specific expression of several specific markers involved in intestinal stem cells, intestinal epithelium, and nutrient absorption. In addition, they showed the key functionality with regard to a high permeability for compounds of up to FITC-dextran 4 kDa, while FITC-dextran 40 kDa failed to enter the organoid lumen and revealed that the genetic properties of bovine intestinal organoids were highly similar to those of in vivo. Collectively, these results provide a reliable method for efficient isolation of intestinal crypts from the small intestine and robust 3D expansion of intestinal organoids in adult bovine and demonstrate the in vitro 3D organoids mimics the in vivo tissue topology and functionality. Finally, intestinal organoids are potential alternatives to in vivo systems and will be facilitated as the practical model to replace animal experiments for various purposes in the fields of animal biotechnology.

Section: Discussionmentioning

confidence: 99%

Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System

Yang

et al. 2021

Animals

“…Recently, intestinal organoids have evolved as potential alternatives to in vivo systems [6,19], and has been a focus of research in livestock species including bovine, swine and chicken [4,5,10,20]. Especially, these organoids has been shown to be an attractive model for mucosal permeability, enabling us to investigate the interactions of pathogenic bacteria, viruses, nutrient absorption and the maintenance of host homeostasis with the gut epithelium of their host [21][22][23][24].…”

Section: Discussionmentioning

confidence: 99%

Robust Three-dimensional (3D) Expansion of Bovine Intestinal Stem Cells: A Model for Replacement of Animal Experiments

Yang

et al. 2021

Preprint

Background Intestinal organoids offer great promise for disease modelling based host-pathogen interactions and nutritional research for feed efficiency measurement in livestock as well as regenerative medicine for therapeutic purposes. However, very limited studies are available on the functional characterization and three-dimensional (3D) expansion of adult stem cells in livestock species compared to mammals. Therefore, we characterized intestinal stem cells derived from small intestine in adult bovine and cultivated intestinal organoids under in vitro 3D culture system.Results In this study, we successfully established intestinal organoids in bovine. Intestinal organoids were long-term cultivated over several passages of culture without loss of the recapitulating capacity of crypts and they had the specific expression of several specific markers involved in intestinal stem cells, intestinal epithelium and nutrient absorption. In addition, they showed the key functionality with regard to a high permeability for compounds of up to FITC-dextran 4 kDa, while FITC-dextran 40 kDa failed to enter the organoid lumen. Furthermore, the genetic properties of intestinal organoids were highly similar to those of in vivo based on QuantSeq 3’ mRNA-Seq. data.Conclusions Collectively, these results provide a reliable method for efficient isolation of intestinal crypts from small intestine and robust 3D expansion of intestinal stem cells in adult bovine and demonstrate the in vitro 3D organoids mimics the in vivo tissue topology and functionality. Finally, intestinal organoids are potential alternatives to in vivo system and will facilitate the practical use of a model to replace animal experiments in the fields of animal biotechnology for various purposes.

“…These intestinal organoids have attracted attention for a variety of applications, including disease modeling, intestinal-microbe interactions, pharmacology, and regenerative medicine, and they have shown promise for the prediction of toxicity ( Bitar and Zakhem, 2016 ; Dedhia et al, 2016 ; Almeqdadi et al, 2019 ). From a toxicological point of view, Belair et al generated 3D spherical ileal organoids from a Matrigel-embedded ileal crypt suspension extracted from commercially available human ileal tissues ( Belair et al, 2020 ). Using ileal organoids, they evaluated 31 marketed drugs known to cause different degrees of clinical incidence of diarrhea with cell viability as a toxicity readout to predict drug-induced diarrhea; they obtained 90% prediction accuracy ( Belair et al, 2020 ).…”

Section: Intestinal Organoids and The Prediction Of Gastrointestinal Toxicitymentioning

confidence: 99%

“…From a toxicological point of view, Belair et al generated 3D spherical ileal organoids from a Matrigel-embedded ileal crypt suspension extracted from commercially available human ileal tissues ( Belair et al, 2020 ). Using ileal organoids, they evaluated 31 marketed drugs known to cause different degrees of clinical incidence of diarrhea with cell viability as a toxicity readout to predict drug-induced diarrhea; they obtained 90% prediction accuracy ( Belair et al, 2020 ). Peters et al developed a non-spherical 3D Epi-Intestinal tissue model by culturing human fibroblasts and primary human small intestinal epithelial cells in collagen conditions in a Transwell format under an air-liquid interface (ALI) ( Peters et al, 2019 ).…”

Section: Intestinal Organoids and The Prediction Of Gastrointestinal Toxicitymentioning

confidence: 99%

“…Since the discovery of human iPSCs and the development of methods for their differentiation into neuronal cells, in vitro predictive assays using iPSC-neurons have been conducted in previous years. Specifically, to predict drug-induced structural and functional toxicity in humans, various readouts such as neuronal cell death, neurite outgrowth, calcium oscillation, and extracellular field potentials were measured in 2D cultured iPSC-derived neurons ( Wilson et al, 2014 ; Bitar and Zakhem, 2016 ; Dedhia et al, 2016 ; Takahashi et al, 2018 ; Almeqdadi et al, 2019 ; Belair et al, 2020 ; Shirakawa and Suzuki, 2020 ).…”

Section: Brain Organoids and The Prediction Of Neurotoxicitymentioning

confidence: 99%

See 1 more Smart Citation

Human Organoids for Predictive Toxicology Research and Drug Development

Matsui

Shinozawa

2021

Front. Genet.

Organoids are three-dimensional structures fabricated in vitro from pluripotent stem cells or adult tissue stem cells via a process of self-organization that results in the formation of organ-specific cell types. Human organoids are expected to mimic complex microenvironments and many of the in vivo physiological functions of relevant tissues, thus filling the translational gap between animals and humans and increasing our understanding of the mechanisms underlying disease and developmental processes. In the last decade, organoid research has attracted increasing attention in areas such as disease modeling, drug development, regenerative medicine, toxicology research, and personalized medicine. In particular, in the field of toxicology, where there are various traditional models, human organoids are expected to blaze a new path in future research by overcoming the current limitations, such as those related to differences in drug responses among species. Here, we discuss the potential usefulness, limitations, and future prospects of human liver, heart, kidney, gut, and brain organoids from the viewpoints of predictive toxicology research and drug development, providing cutting edge information on their fabrication methods and functional characteristics.