Development of in vitro enteroids derived from bovine small intestinal crypts.
Development of in vitro enteroids derived from bovine small intestinal crypts.
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DOI：
10.1186/s13567-018-0547-5
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发表时间：
2018-07-03
影响因子：
4.4
通讯作者：
Mabbott NA
中科院分区：
文献类型：
--
作者：
Hamilton CA;Young R;Jayaraman S;Sehgal A;Paxton E;Thomson S;Katzer F;Hope J;Innes E;Morrison LJ;Mabbott NA
牛是一种重要的经济家畜。肠上皮细胞或上皮细胞系的体外2D培养已广泛用于研究牛肠道中的细胞功能和宿主-病原体相互作用。然而，这些文化缺乏细胞的多样性，在肠上皮细胞中遇到的，和转化细胞系的单培养的生理相关性是不确定的。牛肠上皮细胞分化和稳态的影响因素也知之甚少，很少有细胞特异性标志物，可以区分不同的肠上皮细胞谱系已被报道。在这里，我们描述了一个简单而可靠的程序，建立在体外三维肠，或“小肠”，培养牛小肠（回肠）隐窝。这些肠状体包含一个连续的中央管腔内衬一层极化的肠上皮细胞，其顶端表面上具有丰富的微绒毛的紧密连接结合。组织学和转录分析表明，肠上皮细胞谱系，包括肠干细胞，肠上皮细胞，潘氏细胞，杯状细胞和肠内分泌细胞的混合人口。我们表明，牛肠可以成功地保持长期通过多次连续传代，而没有观察到的变化，其生长特性，形态或转录组。此外，牛类肠可以冷冻保存，并从冷冻储备物中回收活的培养物。我们的数据表明，这些3D牛类肠文化代表了一种新的，生理相关的和易处理的体外系统中，上皮细胞的分化和功能，和宿主-病原体的相互作用，在牛小肠可以进行研究。本文的在线版本（10.1186/s13567-018-0547-5）包含补充材料，可供授权用户使用。
Cattle are an economically important domestic animal species. In vitro 2D cultures of intestinal epithelial cells or epithelial cell lines have been widely used to study cell function and host–pathogen interactions in the bovine intestine. However, these cultures lack the cellular diversity encountered in the intestinal epithelium, and the physiological relevance of monocultures of transformed cell lines is uncertain. Little is also known of the factors that influence cell differentiation and homeostasis in the bovine intestinal epithelium, and few cell-specific markers that can distinguish the different intestinal epithelial cell lineages have been reported. Here we describe a simple and reliable procedure to establish in vitro 3D enteroid, or “mini gut”, cultures from bovine small intestinal (ileal) crypts. These enteroids contained a continuous central lumen lined with a single layer of polarized enterocytes, bound by tight junctions with abundant microvilli on their apical surfaces. Histological and transcriptional analyses suggested that the enteroids comprised a mixed population of intestinal epithelial cell lineages including intestinal stem cells, enterocytes, Paneth cells, goblet cells and enteroendocrine cells. We show that bovine enteroids can be successfully maintained long-term through multiple serial passages without observable changes to their growth characteristics, morphology or transcriptome. Furthermore, the bovine enteroids can be cryopreserved and viable cultures recovered from frozen stocks. Our data suggest that these 3D bovine enteroid cultures represent a novel, physiologically-relevant and tractable in vitro system in which epithelial cell differentiation and function, and host–pathogen interactions in the bovine small intestine can be studied. The online version of this article (10.1186/s13567-018-0547-5) contains supplementary material, which is available to authorized users.
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影响因子：
3.7
作者：
Kuroda K;Kiyono T;Isogai E;Masuda M;Narita M;Okuno K;Koyanagi Y;Fukuda T
通讯作者：
Fukuda T
DOI：
10.1084/jem.20160659
发表时间：
2018-02-05
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Kanaya T;Sakakibara S;Jinnohara T;Hachisuka M;Tachibana N;Hidano S;Kobayashi T;Kimura S;Iwanaga T;Nakagawa T;Katsuno T;Kato N;Akiyama T;Sato T;Williams IR;Ohno H
通讯作者：
Ohno H
影响因子：
3.6
作者：
Khalil HA;Lei NY;Brinkley G;Scott A;Wang J;Kar UK;Jabaji ZB;Lewis M;Martín MG;Dunn JC;Stelzner MG
通讯作者：
Stelzner MG
影响因子：
64.8
作者：
Gerbe F;Sidot E;Smyth DJ;Ohmoto M;Matsumoto I;Dardalhon V;Cesses P;Garnier L;Pouzolles M;Brulin B;Bruschi M;Harcus Y;Zimmermann VS;Taylor N;Maizels RM;Jay P
通讯作者：
Jay P
影响因子：
5.3
作者：
de Lau, Wim;Kujala, Pekka;Clevers, Hans
通讯作者：
Clevers, Hans