Editing Myosin VB Gene to Create Porcine Model of Microvillus Inclusion Disease, With Microvillus-Lined Inclusions and Alterations in Sodium Transporters

Engevik, Amy C.; Coutts, Alexander W.; Kaji, Izumi; Rodriguez, Paula; Ongaratto, Felipe Ledur; Saqui‐Salces, Milena; Medida, Ramya Lekha; Meyer, Anne R.; Kolobova, Elena; Engevik, Melinda A.; Williams, Janice A.; Shub, Mitchell D.; Carlson, Daniel F.; Melkamu, Tamene; Goldenring, James R.

doi:10.1053/j.gastro.2020.02.034

Cited by 27 publications

(39 citation statements)

References 64 publications

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“…The culture of pig, rabbit and bovine intestinal organoid cells in Transwell inserts produced a tight monolayer and the increased transepithelial electrical resistance (TEER) overtime indicated the gradual formation of an intact epithelial barrier [ 19 , 23 ]. In this 2D setting, the apical side of organoid epithelial cells faces up and is thus accessible to experimental treatments [ 16 , 19 , 37 ]. Epithelial differentiation can be induced in 2D monolayer by removing niche factors (as described above) or by using an air liquid interface, as shown for pig enteroids [ 37 ].…”

Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

“…In this 2D setting, the apical side of organoid epithelial cells faces up and is thus accessible to experimental treatments [ 16 , 19 , 37 ]. Epithelial differentiation can be induced in 2D monolayer by removing niche factors (as described above) or by using an air liquid interface, as shown for pig enteroids [ 37 ]. In the latter case, pig organoid cell monolayers were cultured on Transwells for two days in proliferation medium before establishment of the air liquid interface on the third day by removal of the medium in the upper (apical) compartment combined with the switch to differentiation medium lacking Wnt in the lower (basal) compartment [ 37 ].…”

Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

“…Epithelial differentiation can be induced in 2D monolayer by removing niche factors (as described above) or by using an air liquid interface, as shown for pig enteroids [ 37 ]. In the latter case, pig organoid cell monolayers were cultured on Transwells for two days in proliferation medium before establishment of the air liquid interface on the third day by removal of the medium in the upper (apical) compartment combined with the switch to differentiation medium lacking Wnt in the lower (basal) compartment [ 37 ]. The main limitation of monolayers of epithelial cells is the loss of the complex 3D organization of intestinal organoids such as buddings that reproduces in vitro crypt domains.…”

Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

“…Besides its role of barrier, the intestinal epithelium also contributes to food digestion, nutrient absorption and hormonal regulation. Markers of absorptive enterocytes (ALPI, KRT20), digestive enzymes (sucrase-isomaltase), ion/nutrient transporters (MCT1, SGLT1, NHE3) are expressed with the appropriate cellular localization in pig, rabbit, chicken and cow organoids [ 15 , 16 , 19 – 21 , 37 , 39 , 41 ]. Moreover, the characteristic microvilli of mature enterocytes were observed in pig and chicken enteroids [ 19 , 40 ] (Figure 4 F).…”

Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

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Intestinal organoids in farm animals

Beaumont

Blanc

Cherbuy

et al. 2021

Vet Res

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In livestock species, the monolayer of epithelial cells covering the digestive mucosa plays an essential role for nutrition and gut barrier function. However, research on farm animal intestinal epithelium has been hampered by the lack of appropriate in vitro models. Over the past decade, methods to culture livestock intestinal organoids have been developed in pig, bovine, rabbit, horse, sheep and chicken. Gut organoids from farm animals are obtained by seeding tissue-derived intestinal epithelial stem cells in a 3-dimensional culture environment reproducing in vitro the stem cell niche. These organoids can be generated rapidly within days and are formed by a monolayer of polarized epithelial cells containing the diverse differentiated epithelial progeny, recapitulating the original structure and function of the native epithelium. The phenotype of intestinal organoids is stable in long-term culture and reflects characteristics of the digestive segment of origin. Farm animal intestinal organoids can be amplified in vitro, cryopreserved and used for multiple experiments, allowing an efficient reduction of the use of live animals for experimentation. Most of the studies using livestock intestinal organoids were used to investigate host-microbe interactions at the epithelial surface, mainly focused on enteric infections with viruses, bacteria or parasites. Numerous other applications of farm animal intestinal organoids include studies on nutrient absorption, genome editing and bioactive compounds screening relevant for agricultural, veterinary and biomedical sciences. Further improvements of the methods used to culture intestinal organoids from farm animals are required to replicate more closely the intestinal tissue complexity, including the presence of non-epithelial cell types and of the gut microbiota. Harmonization of the methods used to culture livestock intestinal organoids will also be required to increase the reproducibility of the results obtained in these models. In this review, we summarize the methods used to generate and cryopreserve intestinal organoids in farm animals, present their phenotypes and discuss current and future applications of this innovative culture system of the digestive epithelium.

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Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

Section: Phenotype Of Livestock Intestinal Organoidsmentioning

confidence: 99%

See 2 more Smart Citations

Intestinal organoids in farm animals

Beaumont

Blanc

Cherbuy

et al. 2021

Vet Res

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show abstract

“…Recently reported pigs carrying the Navajo MYO5B mutation may therefore be an important tool in development of LPA and other approaches to restoration of membrane traffic to the brush border. 16 The responses of these pigs to LPA will be of great interest.…”

mentioning

confidence: 99%

Exploiting Alternative Brush Border Trafficking Routes to Treat Microvillous Inclusion Disease

Abtahi

Turner

2020

Gastroenterology

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Autosomal recessive diseases among the Athabaskans of the southwestern United States: anthropological, medical, and scientific aspects

Erickson

2021

J Appl Genetics

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The peopling of the Americas by Native Americans occurred in 4 waves of which the last was Nadene language speakers of whom Athabaskans are the largest group. As the Europeans were entering the Southwestern states of the USA, Athabaskan hunting-gathering tribes were migrating South from Canada along the Rocky Mountains and undergoing potential bottlenecks reflected in autosomal recessive diseases shared by Apaches and Navajos. About 300 years ago, the Navajo developing a sedentary culture learned from Pueblo Indians while the Apache remained hunter-gathers. Although most of the tribe was rounded up and forced to relocate to Bosque Redondo, the adult breeding population was large enough to prevent a genetic bottleneck. However, some Navajo underwent further population bottlenecks while hiding from the brutal US Army action (under Kit Carson’s guidance). This led to an increased frequency of other autosomal recessive diseases. Recent advances in population genetics, pathophysiology of the diseases, and social/ethical issues concerning their study are reviewed.

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Editing Myosin VB Gene to Create Porcine Model of Microvillus Inclusion Disease, With Microvillus-Lined Inclusions and Alterations in Sodium Transporters

Cited by 27 publications

References 64 publications

Intestinal organoids in farm animals

Intestinal organoids in farm animals

Exploiting Alternative Brush Border Trafficking Routes to Treat Microvillous Inclusion Disease

Autosomal recessive diseases among the Athabaskans of the southwestern United States: anthropological, medical, and scientific aspects

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