Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine
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 mod...
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doaj-e2c51234984b4abeac4ab23a854b15242020-11-25T03:01:07ZengMDPI AGCells2073-44092020-03-01982282210.3390/cells9040822Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine IntestineNina Kramer0Barbara Pratscher1Andre M. C. Meneses2Waltraud Tschulenk3Ingrid Walter4Alexander Swoboda5Hedwig S. Kruitwagen6Kerstin Schneeberger7Louis C. Penning8Bart Spee9Matthias Kieslinger10Sabine Brandt11Iwan A. Burgener12Division of Small Animal Internal Medicine, Department for Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, AustriaDivision of Small Animal Internal Medicine, Department for Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, AustriaDivision of Small Animal Internal Medicine, Department for Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, AustriaInstitute of Pathology, Department for Pathobiology, University of Veterinary Medicine, 1210 Vienna, AustriaInstitute of Pathology, Department for Pathobiology, University of Veterinary Medicine, 1210 Vienna, AustriaDivision of Small Animal Internal Medicine, Department for Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, AustriaDepartment of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 Utrecht, The NetherlandsDepartment of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 Utrecht, The NetherlandsDepartment of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 Utrecht, The NetherlandsDepartment of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584 Utrecht, The NetherlandsDivision of Small Animal Internal Medicine, Department for Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, AustriaResearch Group Oncology, Equine Surgery, Department of Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, AustriaDivision of Small Animal Internal Medicine, Department for Small Animals and Horses, University of Veterinary Medicine, 1210 Vienna, AustriaFunctional 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.https://www.mdpi.com/2073-4409/9/4/822intestinal organoidscanine intestinedifferentiationorganoid culture |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nina Kramer Barbara Pratscher Andre M. C. Meneses Waltraud Tschulenk Ingrid Walter Alexander Swoboda Hedwig S. Kruitwagen Kerstin Schneeberger Louis C. Penning Bart Spee Matthias Kieslinger Sabine Brandt Iwan A. Burgener |
spellingShingle |
Nina Kramer Barbara Pratscher Andre M. C. Meneses Waltraud Tschulenk Ingrid Walter Alexander Swoboda Hedwig S. Kruitwagen Kerstin Schneeberger Louis C. Penning Bart Spee Matthias Kieslinger Sabine Brandt Iwan A. Burgener Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine Cells intestinal organoids canine intestine differentiation organoid culture |
author_facet |
Nina Kramer Barbara Pratscher Andre M. C. Meneses Waltraud Tschulenk Ingrid Walter Alexander Swoboda Hedwig S. Kruitwagen Kerstin Schneeberger Louis C. Penning Bart Spee Matthias Kieslinger Sabine Brandt Iwan A. Burgener |
author_sort |
Nina Kramer |
title |
Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine |
title_short |
Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine |
title_full |
Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine |
title_fullStr |
Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine |
title_full_unstemmed |
Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine |
title_sort |
generation of differentiating and long-living intestinal organoids reflecting the cellular diversity of canine intestine |
publisher |
MDPI AG |
series |
Cells |
issn |
2073-4409 |
publishDate |
2020-03-01 |
description |
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. |
topic |
intestinal organoids canine intestine differentiation organoid culture |
url |
https://www.mdpi.com/2073-4409/9/4/822 |
work_keys_str_mv |
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