Methionine deficiency and its hydroxy analogue influence chicken intestinal 3-dimensional organoid development.
Methionine deficiency and its hydroxy analogue influence chicken intestinal 3-dimensional organoid development.
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DOI：
10.1016/j.aninu.2021.06.001
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Yuan J
中科院分区：
文献类型：
--
作者：
Wang Y;Hou Q;Wu Y;Xu Y;Liu Y;Chen J;Xu L;Guo Y;Gao S;Yuan J
Methylamine及其羟基类似物（MHA）已被证明有利于小鼠肠道再生。肠道类器官是一个很好的模型，可以直接反映某些营养物质或化学物质对肠道发育的影响。本研究旨在建立一种鸡肠道类器官的体外培养方法，并利用该模型探讨蛋氨酸缺乏和MHA对肠道类器官发育的影响。结果表明，125 μm细胞筛收获鸡胚隐窝的效率最高。我们发现，添加转化生长因子-β抑制剂（A8301）促进肠上皮细胞分化，但牺牲了肠干细胞（ISC）的增殖。丝裂原活化蛋白激酶p38抑制剂（SB 202190）促进肠类器官形成和肠上皮细胞分化，但抑制肠内分泌细胞，杯状细胞和潘氏细胞的分化。然而，在A8301存在下，SB 202190对肠内分泌细胞和潘氏细胞分化的抑制减轻。糖原合成酶激酶3抑制剂（CHIR 99021），丙戊酸（VPA）单独和他们的组合促进鸡肠道类器官的形成和肠上皮细胞分化的潘氏细胞和杯状细胞的表达为代价。鸡血清显著促进类器官的形成，尤其是在A8301，SB 202190，CHIR 99021和VPA的存在下，但抑制潘氏细胞和肠内分泌细胞的分化。0.25%的鸡血清能够满足鸡肠道类器官发育的需要，其对鸡肠道类器官培养的促进作用是胎牛血清和胰岛素样生长因子-1所不能替代的。此外，补充有A8301、SB 202190、CHIR 99021、VPA和鸡血清的商业小鼠类器官培养基促进鸡类器官出芽。基于鸡肠道类器官模型，我们发现，甲硫氨酸缺乏模仿环亮氨酸抑制类器官的形成和类器官的大小，这种效果与环亮氨酸浓度增加加强。与L-蛋氨酸相比，蛋氨酸羟基类似物促进ISC再生，但降低细胞分化。总之，我们的研究结果提供了一个潜在的很好的指导方针，鸡肠道类器官培养和蛋氨酸功能的见解隐窝发展。
Methionine and its hydroxy analogue (MHA) have been shown to benefit mouse intestinal regeneration. The intestinal organoid is a good model that directly reflects the impact of certain nutrients or chemicals on intestinal development. Here, we aimed to establish a chicken intestinal organoid culture method first and then use the model to explore the influence of methionine deficiency and MHA on intestinal organoid development. The results showed that 125-μm cell strainer exhibited the highest efficiency for chicken embryo crypt harvesting. We found that transforming growth factor-β inhibitor (A8301) supplementation promoted enterocyte differentiation at the expense of the proliferation of intestinal stem cells (ISC). The mitogen-activated protein kinase p38 inhibitor (SB202190) promoted intestinal organoid formation and enterocyte differentiation but suppressed the differentiation of enteroendocrine cells, goblet cells and Paneth cells. However, the suppression of enteroendocrine cell and Paneth cell differentiation by SB202190 was alleviated at the presence of A8301. The glycogen synthase kinase 3 inhibitor (CHIR99021), valproic acid (VPA) alone and their combination promoted chicken intestinal organoid formation and enterocyte differentiation at the expense of the expression of Paneth cells and goblet cells. Chicken serum significantly improved organoid formation, especially in the presence of A8301, SB202190, CHIR99021, and VPA, but inhibited the differentiation of Paneth cells and enteroendocrine cells. Chicken serum at a concentration of 0.25% meets the requirement of chicken intestinal organoid development, and the beneficial effect of chicken serum on chicken intestinal organoid culture could not be replaced by fetal bovine serum and insulin-like growth factor-1. Moreover, commercial mouse organoid culture medium supplemented with A8301, SB202190, CHIR99021, VPA, and chicken serum promotes chicken organoid budding. Based on the chicken intestinal organoid model, we found that methionine deficiency mimicked by cycloleucine suppressed organoid formation and organoid size, and this effect was reinforced with increased cycloleucine concentrations. Methionine hydroxy analogue promoted regeneration of ISC but decreased cell differentiation compared with the results obtained with L-methionine. In conclusion, our results provide a potentially excellent guideline for chicken intestinal organoid culture and insights into methionine function in crypt development.
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影响因子：
5.4
作者：
Li Y;Seto E
通讯作者：
Seto E
DOI：
10.1242/dev.163014
发表时间：
2018-05-11
期刊：
Development (Cambridge, England)
影响因子：
--
作者：
Chung MI;Bujnis M;Barkauskas CE;Kobayashi Y;Hogan BLM
通讯作者：
Hogan BLM
影响因子：
4.4
作者：
Li, J.;Li, J., Jr.;Zhang, C. Q.
通讯作者：
Zhang, C. Q.
影响因子：
11.4
作者：
Hoffmann, Karen;Berger, Hilmar;Kessler, Mirjana
通讯作者：
Kessler, Mirjana
影响因子：
2.6
作者：
Acharya, Mohan;Arsi, Komala;Rath, Narayan C.
通讯作者：
Rath, Narayan C.