Probiotic Lactobacillus acidophilus bacteria or synthetic TLR2 agonist boost the growth of chicken embryo intestinal organoids in cultures comprising epithelial cells and myofibroblasts |
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| Institution: | 1. Department of Food Biotechnology, Faculty of Food Technology, The University of Agriculture in Kraków, Balicka 122, 30-149, Kraków, Poland;2. Department of Medicine, Jagiellonian University Medical College, Skawinska 8, 31-066 Kraków, Poland;3. Malopolska Center of Food Monitoring, Faculty of Food Technology, The University of Agriculture in Kraków, Balicka 122, 30-149, Kraków, Poland;1. Department of Medicine, Division of Gastroenterology, Johns Hopkins University School of Medicine, Baltimore, Maryland;2. Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas;3. Department of Gastroenterology and Hepatology, Erasmus University Medical Center, Rotterdam, The Netherlands;1. Koch Institute for Integrative Cancer Research at MIT and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA;2. Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA;1. Center for Cellular Engineering, Clinical Center, National Institutes of Health, Bethesda, MD 20892, USA;2. Translational Stem Cell Biology Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA;3. Angiocrine Bioscience, La Jolla, CA, USA;4. Department of Statistics, University of Washington, Seattle, WA 98195, USA;5. Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 00826, Republic of Korea;6. Flow Cytometry Core, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA;1. Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University, Netherlands;2. Utrecht Medical Center, 3584 CT, Utrecht, Netherlands;2. Statewide Mass spectrometry Facility, Department of Chemistry Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA;3. The Department of Poultry Science, Poultry Science Center, University of Arkansas, Fayetteville, AR 72701, USA |
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| Abstract: | The intestinal epithelial cells reside in close proximity to myofibroblasts and microbiota, which are supposed to have an impact on intestinal stem cells fate and to influence processes of tissue maturation and regeneration. Mechanism underlying these phenomena and their diversity among vertebrates can be studied in 3D organoid cultures. We investigated the growth of chicken embryo intestinal epithelial organoids in Matrigel with and without Toll-like receptors (TLRs) stimulation. The organoid cultures contained also some myofibroblasts with potential to promote intestinal stem cell survival. Organoid cells, expressing TLR4, TLR2 type 1 and TLR2 type 2 were incubated with their agonists (lipopolysaccharide – LPS and Pam3CSK4) or co-cultured with Lactobacillus acidophilus bacteria (LA-5). Pam3CSK4 and LA-5 promoted organoid growth, which was demonstrated by comparing the morphological parameters (mean number and area of organoids). The profile of prostaglandins (PG), known to promote intestinal regeneration, in supernatants from organoid and fibroblast cultures were evaluated. Both PGE2 and PGD2 were detected. As compared to unstimulated controls, supernatants from the Pam3CSK4-stimulated organoids contained twice as much of PGE2 and PGD2. The changes in production of prostaglandins and the support of epithelial cell growth by myofibroblasts are factors potentially responsible for stimulatory effect of TLR2 activation. |
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| Keywords: | Toll-like receptors Intestinal organoids Intestinal myofibroblasts Probiotics Avian immunity |
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