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https://www.readbyqxmd.com/read/27901089/a-silac-based-method-for-quantitative-proteomic-analysis-of-intestinal-organoids
#1
Alexis Gonneaud, Christine Jones, Naomie Turgeon, Dominique Lévesque, Claude Asselin, François Boudreau, François-Michel Boisvert
Organoids have the potential to bridge 3D cell culture to tissue physiology by providing a model resembling in vivo organs. Long-term growing organoids were first isolated from intestinal crypt cells and recreated the renewing intestinal epithelial niche. Since then, this technical breakthrough was applied to many other organs, including prostate, liver, kidney and pancreas. We describe here how to apply a SILAC-based quantitative proteomic approach to measure protein expression changes in intestinal organoids under different experimental conditions...
November 30, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27899814/neurogastroenterology-engineering-human-intestinal-organoids-with-a-functional-ens
#2
Katrina Ray
No abstract text is available yet for this article.
November 30, 2016: Nature Reviews. Gastroenterology & Hepatology
https://www.readbyqxmd.com/read/27889570/inhibitor-of-apoptosis-protein-1-regulates-tumor-necrosis-factor-mediated-destruction-of-intestinal-epithelial-cells
#3
Thomas Grabinger, Konstantin J Bode, Janine Demgenski, Carina Seitz, M Eugenia Delgado, Feodora Kostadinova, Cindy Reinhold, Nima Etemadi, Sabine Wilhelm, Matthias Schweinlin, Kay Hänggi, Janin Knop, Christof Hauck, Heike Walles, John Silke, Harald Wajant, Ueli Nachbur, W Wei-Lynn Wong, Thomas Brunner
BACKGROUND AND AIMS: Tumor necrosis factor (TNF) is a cytokine that promotes inflammation and contributes to pathogenesis of inflammatory bowel diseases. Unlike other cells and tissues, intestinal epithelial cells undergo rapid cell death upon exposure to TNF, by unclear mechanisms. We investigated the roles of inhibitor of apoptosis proteins (IAPs) in the regulation of TNF-induced cell death in the intestinal epithelium of mice and intestinal organoids. METHODS: RNA from cell lines and tissues and analyzed by quantitative PCR, protein levels were analyzed by immunoblot assays...
November 23, 2016: Gastroenterology
https://www.readbyqxmd.com/read/27888356/intestinal-stem-cell-transplantation
#4
REVIEW
Tetsuya Nakamura, Mamoru Watanabe
Organoid technologies to expand intestinal epithelial cells are gaining increasing attention as a useful tool to investigate many aspects of intestinal epithelial biology and pathology. One important application of organoid systems would be to use intestinal epithelial cells expanded in culture for following transplantation experiments. In this article, we present a brief overview of the studies that have succeeded in generating new epithelial tissues in the surface of native intestines in mice by organoid transplantation...
November 25, 2016: Journal of Gastroenterology
https://www.readbyqxmd.com/read/27882268/long-term-culture-of-intestinal-cell-progenitors-an-overview-of-their-development-application-and-associated-technologies
#5
REVIEW
Andrew J Hollins, Lee Parry
PURPOSE OF REVIEW: Long-term culture of adult progenitor cells in 3D is a recently emerging technology that inhabits the space between 2D cell lines and organ slice culture. RECENT FINDINGS: Adaptations to defined media components in the wake of advances in ES and iPS cell culture has led to the identification of conditions that maintained intestinal cell progenitors in culture. These conditions retain cellular heterogeneity of the normal or tumour tissue, and the cultures have been shown to be genetically stable, such that substantial biobanks are being created from patient derived material...
2016: Current Pathobiology Reports
https://www.readbyqxmd.com/read/27869805/engineered-human-pluripotent-stem-cell-derived-intestinal-tissues-with-a-functional-enteric-nervous-system
#6
Michael J Workman, Maxime M Mahe, Stephen Trisno, Holly M Poling, Carey L Watson, Nambirajan Sundaram, Ching-Fang Chang, Jacqueline Schiesser, Philippe Aubert, Edouard G Stanley, Andrew G Elefanty, Yuichiro Miyaoka, Mohammad A Mandegar, Bruce R Conklin, Michel Neunlist, Samantha A Brugmann, Michael A Helmrath, James M Wells
The enteric nervous system (ENS) of the gastrointestinal tract controls many diverse functions, including motility and epithelial permeability. Perturbations in ENS development or function are common, yet there is no human model for studying ENS-intestinal biology and disease. We used a tissue-engineering approach with embryonic and induced pluripotent stem cells (PSCs) to generate human intestinal tissue containing a functional ENS. We recapitulated normal intestinal ENS development by combining human-PSC-derived neural crest cells (NCCs) and developing human intestinal organoids (HIOs)...
November 21, 2016: Nature Medicine
https://www.readbyqxmd.com/read/27869785/classic-il-6r-signalling-is-dispensable-for-intestinal-epithelial-proliferation-and-repair
#7
K Aden, A Breuer, A Rehman, H Geese, F Tran, J Sommer, G H Waetzig, T M Reinheimer, S Schreiber, S Rose-John, J Scheller, P Rosenstiel
Inflammatory bowel disease is characterized by disturbed cytokine signalling in the mucosa. Inhibition of the proinflammatory interleukin (IL)-6 pathway is a promising new therapeutic strategy, but safety concerns arise as IL-6 signalling also contributes to epithelial repair of the intestinal mucosa. To which extent IL-6 classic or trans-signalling contributes to intestinal repair remains elusive. We tested the influence of IL-6 classic signalling on intestinal repair and proliferation. Whereas IL-6 induced STAT3 phosphorylation in the colonic cancer cell lines, primary non-malignant intestinal organoids did not respond to IL-6 classic signalling...
November 21, 2016: Oncogenesis
https://www.readbyqxmd.com/read/27867006/intercellular-coupling-of-the-cell-cycle-and-circadian-clock-in-adult-stem-cell-culture
#8
Toru Matsu-Ura, Andrey Dovzhenok, Eitaro Aihara, Jill Rood, Hung Le, Yan Ren, Andrew E Rosselot, Tongli Zhang, Choogon Lee, Karl Obrietan, Marshall H Montrose, Sookkyung Lim, Sean R Moore, Christian I Hong
Circadian clock-gated cell division cycles are observed from cyanobacteria to mammals via intracellular molecular connections between these two oscillators. Here we demonstrate WNT-mediated intercellular coupling between the cell cycle and circadian clock in 3D murine intestinal organoids (enteroids). The circadian clock gates a population of cells with heterogeneous cell-cycle times that emerge as 12-hr synchronized cell division cycles. Remarkably, we observe reduced-amplitude oscillations of circadian rhythms in intestinal stem cells and progenitor cells, indicating an intercellular signal arising from differentiated cells governing circadian clock-dependent synchronized cell division cycles...
December 1, 2016: Molecular Cell
https://www.readbyqxmd.com/read/27856615/epigenetic-regulation-of-intestinal-stem-cells-by-tet1-mediated-dna-hydroxymethylation
#9
Rinho Kim, Karyn L Sheaffer, Inchan Choi, Kyoung-Jae Won, Klaus H Kaestner
Methylated cytosines are associated with gene silencing. The ten-eleven translocation (TET) hydroxylases, which oxidize methylated cytosines to 5-hydroxymethylcytosine (5hmC), are essential for cytosine demethylation. Gene silencing and activation are critical for intestinal stem cell (ISC) maintenance and differentiation, but the potential role of TET hydroxylases in these processes has not yet been examined. Here, we generated genome-wide maps of the 5hmC mark in ISCs and their differentiated progeny. Genes with high levels of hydroxymethylation in ISCs are strongly associated with Wnt signaling and developmental processes...
November 17, 2016: Genes & Development
https://www.readbyqxmd.com/read/27856416/regulation-and-function-of-bone-morphogenetic-protein-signaling-in-colonic-injury-and-inflammation
#10
Tuo Ji, Hidehiko Takabayashi, Maria Mao, Xu Han, Xiang Xue, Jennifer C Brazil, Kathryn A Eaton, Yatrik M Shah, Andrea Todisco
The bone morphogenetic proteins (BMPs) regulate gastrointestinal homeostasis. We investigated the expression of BMP-4 and the localization and function of BMP signaling during colonic injury and inflammation. Wild type mice and mice expressing the β-galactosidase (β-gal) gene under the control of a BMP responsive element (BRE), BMP-4-β-gal/+ mice, and animals generated by crossing villin-Cre mice to mice with floxed alleles of BMP receptor 1A (villin-Cre;Bmpr1a(flox/flox)), were treated with DSS to induce colonic injury and inflammation...
November 17, 2016: American Journal of Physiology. Gastrointestinal and Liver Physiology
https://www.readbyqxmd.com/read/27851739/designer-matrices-for-intestinal-stem-cell-and-organoid-culture
#11
Nikolce Gjorevski, Norman Sachs, Andrea Manfrin, Sonja Giger, Maiia E Bragina, Paloma Ordóñez-Morán, Hans Clevers, Matthias P Lutolf
Epithelial organoids recapitulate multiple aspects of real organs, making them promising models of organ development, function and disease. However, the full potential of organoids in research and therapy has remained unrealized, owing to the poorly defined animal-derived matrices in which they are grown. Here we used modular synthetic hydrogel networks to define the key extracellular matrix (ECM) parameters that govern intestinal stem cell (ISC) expansion and organoid formation, and show that separate stages of the process require different mechanical environments and ECM components...
November 16, 2016: Nature
https://www.readbyqxmd.com/read/27827428/pge2-is-a-direct-and-robust-mediator-of-anion-fluid-secretion-by-human-intestinal-epithelial-cells
#12
Satoru Fujii, Kohei Suzuki, Ami Kawamoto, Fumiaki Ishibashi, Toru Nakata, Tatsuro Murano, Go Ito, Hiromichi Shimizu, Tomohiro Mizutani, Shigeru Oshima, Kiichiro Tsuchiya, Tetsuya Nakamura, Akihiro Araki, Kazuo Ohtsuka, Ryuichi Okamoto, Mamoru Watanabe
Intestinal epithelial cells (IECs) play an indispensable role in maintaining body fluid balance partly through their ability to regulate anion/fluid secretion. Yet in various inflammatory gastrointestinal diseases, over-secretion of anions results in symptoms such as severe diarrhoea. Endogenous mediators, such as vasoactive intestinal peptide or prostaglandin E2 (PGE2), regulate intestinal anion/fluid secretion, but their direct effect on purified human IECs has never been described in detail. Based on a previously described intestinal organoid swelling model, we established a 3D-scanner-assisted quantification method to evaluate the anion/fluid secretory response of cultured human IECs...
November 9, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27819675/induction-of-endoplasmic-reticulum-stress-by-deletion-of-grp78-depletes-apc-mutant-intestinal-epithelial-stem-cells
#13
J F van Lidth de Jeude, B J Meijer, M C B Wielenga, C N Spaan, B Baan, S L Rosekrans, S Meisner, Y H Shen, A S Lee, J C Paton, A W Paton, V Muncan, G R van den Brink, J Heijmans
Intestinal epithelial stem cells are highly sensitive to differentiation induced by endoplasmic reticulum (ER) stress. Colorectal cancer develops from mutated intestinal epithelial stem cells. The most frequent initiating mutation occurs in Apc, which results in hyperactivated Wnt signalling. This causes hyperproliferation and reduced sensitivity to chemotherapy, but whether these mutated stem cells are sensitive to ER stress induced differentiation remains unknown. Here we examined this by generating mice in which both Apc and ER stress repressor chaperone Grp78 can be conditionally deleted from the intestinal epithelium...
November 7, 2016: Oncogene
https://www.readbyqxmd.com/read/27803115/alterations-in-the-epithelial-stem-cell-compartment-could-contribute-to-permanent-changes-in-the-mucosa-of-patients-with-ulcerative-colitis
#14
Isabella Dotti, Rut Mora-Buch, Elena Ferrer-Picón, Núria Planell, Peter Jung, M Carme Masamunt, Raquel Franco Leal, Javier Martín de Carpi, Josep Llach, Ingrid Ordás, Eduard Batlle, Julián Panés, Azucena Salas
OBJECTIVE: UC is a chronic inflammatory disease of the colonic mucosa. Growing evidence supports a role for epithelial cell defects in driving pathology. Moreover, long-lasting changes in the epithelial barrier have been reported in quiescent UC. Our aim was to investigate whether epithelial cell defects could originate from changes in the epithelial compartment imprinted by the disease. DESIGN: Epithelial organoid cultures (EpOCs) were expanded ex vivo from the intestinal crypts of non-IBD controls and patients with UC...
November 1, 2016: Gut
https://www.readbyqxmd.com/read/27798332/the-noncaloric-sweetener-rebaudioside-a-stimulates-glucagon-like-peptide-1-release-and-increases-enteroendocrine-cell-numbers-in-2-dimensional-mouse-organoids-derived-from-different-locations-of-the-intestine
#15
Nikkie van der Wielen, Jean Paul Ten Klooster, Susanne Muckenschnabl, Raymond Pieters, Henk Fj Hendriks, Renger F Witkamp, Jocelijn Meijerink
BACKGROUND: Glucagon-like peptide 1 (GLP-1) contributes to satiety and plays a pivotal role in insulin secretion and glucose homeostasis. Similar to GLP-1, peptide YY (PYY) and cholecystokinin also influence food intake. The secretion of these hormones by enteroendocrine cells along the intestine is modulated by nutrients. Preparations from the Stevia rebaudiana plant, including rebaudioside A, are increasingly being used as noncaloric sweeteners. OBJECTIVE: We investigated the effects of rebaudioside A on enteroendocrine cells by assessing both cell numbers as well as their secretory capacity in an organoid model...
October 26, 2016: Journal of Nutrition
https://www.readbyqxmd.com/read/27787775/the-three-dimensional-culture-of-epithelial-organoids-derived-from-embryonic-chicken-intestine
#16
Malgorzata Pierzchalska, Malgorzata Panek, Malgorzata Czyrnek, Maja Grabacka
The intestinal epithelium isolated from chicken embryos in last 3 days of development can be used to establish the 3D culture of intestinal organoids. When fragments of epithelial tissue released by incubation with EGTA (2.5 mM, 2 h) are embedded in Matrigel matrix on cell culture inserts the formation of empty spheres covered by epithelial cells is observed in first 24 h of culture. The growth and survival of organoids are supported by the addition of R-spondin 1, Noggin, and prostaglandin E2 to the culture medium...
October 28, 2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27729384/long-term-inflammation-transforms-intestinal-epithelial-cells-of-colonic-organoids
#17
Shuji Hibiya, Kiichiro Tsuchiya, Ryohei Hayashi, Keita Fukushima, Nobukatsu Horita, Sho Watanabe, Tomoaki Shirasaki, Ryu Nishimura, Natsuko Kimura, Tatsunori Nishimura, Noriko Gotoh, Shigeru Oshima, Ryuichi Okamoto, Tetsuya Nakamura, Mamoru Watanabe
BACKGROUND & AIMS: Patients with ulcerative colitis (UC) are at an increased risk of developing colitis-associated cancer (CAC), suggesting that continuous inflammation in the colon promotes the transformation of colonic epithelial cells. However, the mechanisms underlying cell transformation in UC remain unknown. We therefore aimed to investigate the effect of long-term inflammation on intestinal epithelial cells (IECs) using organoid culture. METHODS: IECs were isolated from mice colon, and were cultured according to a method for a three-dimensional (3D) organoid culture...
October 11, 2016: Journal of Crohn's & Colitis
https://www.readbyqxmd.com/read/27713700/drug-discovery-via-human-derived-stem-cell-organoids
#18
Fangkun Liu, Jing Huang, Bo Ning, Zhixiong Liu, Shen Chen, Wei Zhao
Patient-derived cell lines and animal models have proven invaluable for the understanding of human intestinal diseases and for drug development although both inherently comprise disadvantages and caveats. Many genetically determined intestinal diseases occur in specific tissue microenvironments that are not adequately modeled by monolayer cell culture. Likewise, animal models incompletely recapitulate the complex pathologies of intestinal diseases of humans and fall short in predicting the effects of candidate drugs...
2016: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/27704362/isolation-and-culture-of-adult-intestinal-gastric-and-liver-organoids-for-cre-recombinase-mediated-gene-deletion
#19
Dustin J Flanagan, Renate H M Schwab, Bang M Tran, Toby J Phesse, Elizabeth Vincan
The discovery of Lgr5 as a marker of adult stem cells meant that stem cell populations could be purified and studied in isolation. Importantly, when cultured under the appropriate conditions these stem cells form organoids in tissue culture that retain many features of the tissue of origin. The organoid cultures are accessible to genetic and biochemical manipulation, bridging the gap between in vivo mouse models and conventional tissue culture. Here we describe robust protocols to establish organoids from gastrointestinal tissues (stomach, intestine, liver) and Cre-recombinase mediated gene manipulation in vitro...
October 5, 2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27698416/tissue-specific-mutation-accumulation-in-human-adult-stem-cells-during-life
#20
Francis Blokzijl, Joep de Ligt, Myrthe Jager, Valentina Sasselli, Sophie Roerink, Nobuo Sasaki, Meritxell Huch, Sander Boymans, Ewart Kuijk, Pjotr Prins, Isaac J Nijman, Inigo Martincorena, Michal Mokry, Caroline L Wiegerinck, Sabine Middendorp, Toshiro Sato, Gerald Schwank, Edward E S Nieuwenhuis, Monique M A Verstegen, Luc J W van der Laan, Jeroen de Jonge, Jan N M IJzermans, Robert G Vries, Marc van de Wetering, Michael R Stratton, Hans Clevers, Edwin Cuppen, Ruben van Boxtel
The gradual accumulation of genetic mutations in human adult stem cells (ASCs) during life is associated with various age-related diseases, including cancer. Extreme variation in cancer risk across tissues was recently proposed to depend on the lifetime number of ASC divisions, owing to unavoidable random mutations that arise during DNA replication. However, the rates and patterns of mutations in normal ASCs remain unknown. Here we determine genome-wide mutation patterns in ASCs of the small intestine, colon and liver of human donors with ages ranging from 3 to 87 years by sequencing clonal organoid cultures derived from primary multipotent cells...
October 3, 2016: Nature
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