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一种在生理氧含量下培养人类肠道类器官的新系统,用于研究微生物-宿主相互作用。

A novel system to culture human intestinal organoids under physiological oxygen content to study microbial-host interaction.

机构信息

Alkek Centre for Metagenomics and Microbiome Research, Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, United States of America.

Department of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, NE, United States of America.

出版信息

PLoS One. 2024 Jul 25;19(7):e0300666. doi: 10.1371/journal.pone.0300666. eCollection 2024.

Abstract

Mechanistic investigation of host-microbe interactions in the human gut are hindered by difficulty of co-culturing microbes with intestinal epithelial cells. On one hand the gut bacteria are a mix of facultative, aerotolerant or obligate anaerobes, while the intestinal epithelium requires oxygen for growth and function. Thus, a coculture system that can recreate these contrasting oxygen requirements is critical step towards our understanding microbial-host interactions in the human gut. Here, we demonstrate Intestinal Organoid Physoxic Coculture (IOPC) system, a simple and cost-effective method for coculturing anaerobic intestinal bacteria with human intestinal organoids (HIOs). Using commensal anaerobes with varying degrees of oxygen tolerance, such as nano-aerobe Bacteroides thetaiotaomicron and strict anaerobe Blautia sp., we demonstrate that IOPC can successfully support 24-48 hours HIO-microbe coculture. The IOPC recapitulates the contrasting oxygen conditions across the intestinal epithelium seen in vivo. The IOPC cultured HIOs showed increased barrier integrity, and induced expression of immunomodulatory genes. A transcriptomic analysis suggests that HIOs from different donors show differences in the magnitude of their response to coculture with anaerobic bacteria. Thus, the IOPC system provides a robust coculture setup for investigating host-microbe interactions in complex, patient-derived intestinal tissues, that can facilitate the study of mechanisms underlying the role of the microbiome in health and disease.

摘要

研究人体肠道中宿主-微生物相互作用的机制受到与肠道上皮细胞共培养微生物的困难所阻碍。一方面,肠道细菌是兼性需氧菌、耐氧厌氧菌或专性厌氧菌的混合物,而肠道上皮细胞的生长和功能需要氧气。因此,能够再现这些对比氧需求的共培养系统是我们理解人类肠道中微生物-宿主相互作用的关键步骤。在这里,我们展示了肠道类器官低氧共培养(IOPC)系统,这是一种简单且具有成本效益的方法,用于将厌氧肠道细菌与人类肠道类器官(HIOs)共培养。使用具有不同程度耐氧性的共生厌氧菌,如纳米需氧菌拟杆菌和严格厌氧菌布劳特氏菌,我们证明 IOPC 可以成功地支持 24-48 小时的 HIO-微生物共培养。IOPC 再现了体内观察到的肠道上皮细胞中对比鲜明的氧气条件。IOPC 培养的 HIOs 显示出增加的屏障完整性,并诱导免疫调节基因的表达。转录组分析表明,来自不同供体的 HIOs 对与厌氧菌共培养的反应程度存在差异。因此,IOPC 系统为研究复杂的、源自患者的肠道组织中的宿主-微生物相互作用提供了一个强大的共培养设置,这有助于研究微生物组在健康和疾病中的作用的机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ed1a/11271918/62d26a13dc3d/pone.0300666.g001.jpg

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