Project description:RNA sequencing was performed for the TNF-free and -treated enteroids derived from control (n=8 pairs) and Crohn's disease patients (n=8 pairs).
Project description:Organoids grown from intestinal crypts of controls and patients with Crohn's disease were sub-cultured at least 6 passages. RNA sequencing was performed for the 12 paired TNF-free and -treated control and Crohn's organoids.
Project description:Organoids grown from intestinal crypts of controls and patients with Crohn's disease were sub-cultured at least 6 passages. Single cell RNA sequencing was performed for the paired TNF-free and -treated control and Crohn's organoids.
Project description:Intestinal tissues from 3 infants and 3 adults were obtained and the stem cells were isolated to generate enteroids. Enteroids were differentiated for 5 days prior to RNA extraction and sequencing.
Project description:The goal of this study was to evaluate the transcriptional response of human enteroids/colonoids on transwells to infections (bacterial and rotavirus). Enteroids/colonoids lines C103, C109, D103, D109, I103, I109, J2 and J11 were plated on transwells coated with Matrigel, differentiated, and inoculated (rotavirus (Ito), bacteria or mock) for 6 or 24 hours. Subsequently, total RNA was isolated and paired-end sequencing was performed.
Project description:To investigate the effect of human norovirus infection on the trascriptome of intestinal enteroids in the presence of a Jak kinase inhibitor, Ruxolitinib
Project description:The goal of this experiment is to decipher the mechanism of intestinal endocrine / enteroendocrine subtype specification, that is far from being fully understood. To this end, we used single cell genomics in mouse mini-guts. Enteroendocrine cells are derived from endocrine progenitors expressing the transcription factor Ngn3. We took advantage of the Ngn3+/eYFP mouse model -where endocrine progenitors and their descendants can be isolated and sorted by FACS on the basis of the eYFP fluorescence- and established enteroids or mini-guts from the small intestine. Enteroids were dissociated and eYFP+ single cells were directly sorted in 96 wells of the Precise WTA Single Cell Encoding Plate (BD™ Precise WTA Single Cell Kit, BD Genomics). cDNA and libraries were prepared following the BD protocol. Sequencing (paired-end, 2x100b) was performed in a HiSeq 4000 (Illumina). The bioinformatic analysis allowed to identify 8 different groups of enteroendocrine cells with specific signatures.
Project description:In vitro 2D cultures of intestinal epithelial cells or epithelial cell lines have been widely used to study cell function and host-pathogen interactions in the bovine intestine. However, these cultures lack the cellular diversity encountered in the intestinal epithelium, and the physiological relevance of monocultures of transformed cell lines is uncertain. Little is also known of the factors that influence cell differentiation and homeostasis in the bovine intestinal epithelium, and few cell-specific markers that can distinguish the different intestinal epithelial cell lineages have been reported. Here we describe a simple and reliable procedure to establish in vitro 3D enteroid, or “mini gut”, cultures from bovine small intestinal (ileal) crypts. These enteroids contained a continuous central lumen lined with a single layer of polarized enterocytes, bound by tight junctions with abundant microvilli on their apical surfaces. Histological and transcriptional analyses suggested that the enteroids comprised a mixed population of intestinal epithelial cell lineages including intestinal stem cells, enterocytes, Paneth cells, goblet cells and enteroendocrine cells. We show that bovine enteroids can be successfully maintained long-term through multiple serial passages without observable changes to their growth characteristics, morphology or transcriptome. Furthermore, the bovine enteroids can be cryopreserved and viable cultures recovered from frozen stocks. Our data suggest that these 3D bovine enteroid cultures represent a novel, physiologically-relevant and tractable in vitro system in which epithelial cell differentiation and function, and host-pathogen interactions in the bovine small intestine can be studied.