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Open AccessArticle

Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine

1
Division of Small Animal Internal Medicine, Department for Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, Austria
2
Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 Utrecht, The Netherlands
3
Institute of Pathology, Department for Pathobiology, University of Veterinary Medicine, 1210 Vienna, Austria
4
Research Group Oncology, Equine Surgery, Department of Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, Austria
*
Author to whom correspondence should be addressed.
Current address: Institute of Animal Health and Production, Veterinary Hospital, Amazon Rural Federal University, Belém 66.077-830, Brazil.
These authors contributed equally to this work.
Cells 2020, 9(4), 822; https://doi.org/10.3390/cells9040822
Received: 17 February 2020 / Revised: 23 March 2020 / Accepted: 26 March 2020 / Published: 28 March 2020
(This article belongs to the Special Issue 3D Stem Cell Culture)
Functional intestinal disorders constitute major, potentially lethal health problems in humans. Consequently, research focuses on elucidating the underlying pathobiological mechanisms and establishing therapeutic strategies. In this context, intestinal organoids have emerged as a potent in vitro model as they faithfully recapitulate the structure and function of the intestinal segment they represent. Interestingly, human-like intestinal diseases also affect dogs, making canine intestinal organoids a promising tool for canine and comparative research. Therefore, we generated organoids from canine duodenum, jejunum and colon, and focused on simultaneous long-term expansion and cell differentiation to maximize applicability. Following their establishment, canine intestinal organoids were grown under various culture conditions and then analyzed with respect to cell viability/apoptosis and multi-lineage differentiation by transcription profiling, proliferation assay, cell staining, and transmission electron microscopy. Standard expansion medium supported long-term expansion of organoids irrespective of their origin, but inhibited cell differentiation. Conversely, transfer of organoids to differentiation medium promoted goblet cell and enteroendocrine cell development, but simultaneously induced apoptosis. Unimpeded stem cell renewal and concurrent differentiation was achieved by culturing organoids in the presence of tyrosine kinase ligands. Our findings unambiguously highlight the characteristic cellular diversity of canine duodenum, jejunum and colon as fundamental prerequisite for accurate in vitro modelling. View Full-Text
Keywords: intestinal organoids; canine intestine; differentiation; organoid culture intestinal organoids; canine intestine; differentiation; organoid culture
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MDPI and ACS Style

Kramer, N.; Pratscher, B.; Meneses, A.M.C.; Tschulenk, W.; Walter, I.; Swoboda, A.; Kruitwagen, H.S.; Schneeberger, K.; Penning, L.C.; Spee, B.; Kieslinger, M.; Brandt, S.; Burgener, I.A. Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine. Cells 2020, 9, 822. https://doi.org/10.3390/cells9040822

AMA Style

Kramer N, Pratscher B, Meneses AMC, Tschulenk W, Walter I, Swoboda A, Kruitwagen HS, Schneeberger K, Penning LC, Spee B, Kieslinger M, Brandt S, Burgener IA. Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine. Cells. 2020; 9(4):822. https://doi.org/10.3390/cells9040822

Chicago/Turabian Style

Kramer, Nina; Pratscher, Barbara; Meneses, Andre M.C.; Tschulenk, Waltraud; Walter, Ingrid; Swoboda, Alexander; Kruitwagen, Hedwig S.; Schneeberger, Kerstin; Penning, Louis C.; Spee, Bart; Kieslinger, Matthias; Brandt, Sabine; Burgener, Iwan A. 2020. "Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine" Cells 9, no. 4: 822. https://doi.org/10.3390/cells9040822

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