Phorbol 12-Myristate 13-Acetate-Induced Changes in Chicken Enterocytes

et al. 2020

Preprint

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Background: Three-dimensional models of cell culture such as organoids and mini organs accord better advantage over regular cell culture because of their ability to simulate organ functions hence, used for disease modeling, metabolic research, and the development of therapeutics strategies. However, most advances in this area are limited to mammalian species with little progress in others such as poultry where it can be deployed to study problems of agricultural importance. In the course of enterocyte culture in chicken, we observed that intestinal mucosal villus-crypts self-repair and form spheroid-like structures which appear to be useful as ex vivo models to study enteric physiology and diseases. Results: The villus-crypts harvested from chicken intestinal mucosa were cultured to generate enteroids, purified by filtration then re cultured with different chemicals and growth factors to assess their response based on their morphological dispositions. Histochemical analyses using marker antibodies and probes showed the enteroids consisting different cell types such as epithelial, goblet, and enteroendocrine cells typical to villi and retain functional characteristics of intestinal mucosa.Conclusions: We present a simple procedure to generate avian crypt-villous enteroids containing different cell types. Because the absorptive cells are functionally positioned outwards, similar to the luminal enterocytes, the cells have better advantages to interact with the factors present in the culture medium. Thus, the enteroids have the potential to study the physiology, metabolism, and pathology of the intestinal villi and can be useful for preliminary screenings of the factors that may affect gut health in a cost-effective manner and reduce the use of live animals.

Section: Discussionsupporting

confidence: 77%

Production and characterization of avian crypt-villus enteroids and the effect of chemicals

et al. 2020

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“…In the current study, indomethacin appeared to affect the outer epithelial cells of the enteroids without causing any lethality. In a previous study using PMA, we observed significant cachectic effect of it on the enterocytes at concentrations of 1µg/ml or less [23,24], however, it showed no significant effect on the enteroids. Monensin, an anticoccidial drug used in poultry production caused significant damage to the organoids and some early studies it was reported to have toxic and necrotic effect on intestine [73].…”

Section: Discussionmentioning

confidence: 68%

Production and the characterization of avian crypt-villus enteroids and the effect of chemicals

et al. 2020

Preprint

Self Cite

Background: Three-dimensional models of cell culture such as organoids and mini organs accord better advantage over regular cell culture because of their ability to simulate organ functions hence, used for disease modeling, metabolic research, and the development of therapeutics strategies. However, most advances in this area are limited to mammalian species with little progress in others such as poultry where it can be deployed to study problems of agricultural importance. In the course of enterocyte culture in chicken, we observed that intestinal mucosal villus-crypts self-repair and form spheroid-like structures which appear to be useful as ex vivo models to study enteric physiology and diseases. Results: The villus-crypts harvested from chicken intestinal mucosa were cultured to generate enteroids, purified by filtration then re cultured with different chemicals and growth factors to assess their response based on their morphological dispositions. Histochemical analyses using marker antibodies and probes showed the enteroids consisting different cell types such as epithelial, goblet, and enteroendocrine cells typical to villi and retain functional characteristics of intestinal mucosa. Conclusions: We present a simple procedure to generate avian crypt-villous enteroids containing different cell types. Because the absorptive cells are functionally positioned outwards, similar to the luminal enterocytes, the cells have better advantages to interact with the factors present in the culture medium. Thus, the enteroids have the potential to study the physiology, metabolism, and pathology of the intestinal villi and can be useful for preliminary screenings of the factors that may affect gut health in a cost-effective manner and reduce the use of live animals.

“…The effect of some metabolic modulators, indomethacin, a prostaglandin inhibitor, PMA, a protein kinase C activator, and the monensin, an ionophore, anticoccidial antibiotic, on the enteroid morphologies were tested. Indomethacin produces ulcers in human and rat small intestines [72] however, it appeared only to shrink epithelial aspects of the enteroids whereas PMA showed no significant effect on the enteroids although our previous study with the effect of PMA on enterocytes showed significant cachectic changes in the enterocytes [23,24]. Monensin, on the other hand, damaged the organoids, however, it has been a preferred anticoccidial drug in the poultry and some early studies suggested its toxic and necrotic effect in the intestine [73].…”

Section: Discussionmentioning

confidence: 94%

“…Large scale screening assays may be possible if the organoids can be generated rapidly and cost-effectively. In the course of developing chicken enterocyte culture [23,24], we observed that the villus crypts of intestinal mucosa tend to self-repair to form spheroid-like structures resembling mini organoids that may have potential to study villus physiology. Hence, the objective of this study was to streamline the procedure, purify avian villus enteroids, characterize them, and study their potential as a screening tool using some selective chemicals and growth factors.…”

mentioning

confidence: 99%

Production and the characterization of avian crypt-villus enteroids and the effect of chemicals

et al. 2019

Preprint

Self Cite

Background: Three-dimensional models of cell culture such as organoids and mini organs accord better advantage over regular cell culture because of their ability to simulate organ functions hence, used for disease modeling, metabolic research, and the development of therapeutics strategies. However, most advances in this area are limited to mammalian species with little progress in others such as poultry where it can be deployed to study problems of agricultural importance. In the course of enterocyte culture in chicken, we observed that intestinal mucosal villus-crypts self-repair and lead to the formation of enteroid-like structures which appeared to be useful as ex vivo models to study enteric physiology and diseases. Results: The villus-crypts harvested from chicken intestinal mucosa were cultured to generate enteroids, purified by filtration then re cultured with different chemicals and growth factors to assess their response based on their morphological dispositions. Histochemical analyses using marker antibodies and probes showed the enteroids consisting different cells such as epithelial, goblet, and enteroendocrine cells typical to villi and retain functional characteristics of intestinal mucosa. Conclusions: The villus enteroids can simulate villus functions with their absorptive cells functionally positioned and exposed to culture medium thus, can help understand the mechanisms of nutrient uptake and their regulation, interactions with alien organisms, and amenable to assays of the factors that may affect gut health.