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Identification, isolation and analysis of human gut-associated lymphoid tissues
Nature Protocols ( IF 14.8 ) Pub Date : 2021-02-22 , DOI: 10.1038/s41596-020-00482-1
Peter B. Jørgensen , Thomas M. Fenton , Urs M. Mörbe , Lene B. Riis , Henrik L. Jakobsen , Ole H. Nielsen , William W. Agace

Gut-associated lymphoid tissues (GALTs) comprise key intestinal immune inductive sites, including the Peyer’s patches of the small intestine and different types of isolated lymphoid follicle (ILF) found along the length of the gut. Our understanding of human GALT is limited due to a lack of protocols for their isolation. Here we describe a technique that, uniquely among intestinal cell isolation protocols, allows identification and isolation of all human GALT, as well as GALT-free intestinal lamina propria (LP). The technique involves the mechanical separation of intestinal mucosa from the submucosa, allowing the identification and isolation of submucosal ILF (SM-ILF), LP-embedded mucosal ILF (M-ILF) and LP free of contaminating lymphoid tissue. Individual SM-ILF, M-ILF and Peyer’s patch follicles can be subsequently digested for downstream cellular and molecular characterization. The technique, which takes 4–10 h, will be useful for researchers interested in intestinal immune development and function in health and disease.



中文翻译:

人类肠道相关淋巴组织的鉴定、分离和分析

肠道相关淋巴组织 (GALT) 包括关键的肠道免疫诱导位点,包括小肠的派尔斑和沿肠道长度发现的不同类型的孤立淋巴滤泡 (ILF)。由于缺乏隔离协议,我们对人类 GALT 的了解有限。在这里,我们描述了一种技术,该技术在肠道细胞分离方案中独一无二,允许识别和分离所有人类 GALT,以及无 GALT 的肠道固有层 (LP)。该技术涉及从黏膜下层机械分离肠黏膜,从而可以识别和分离黏膜下层 ILF (SM-ILF)、LP 包埋黏膜 ILF (M-ILF) 和无污染淋巴组织的 LP。个人 SM-ILF,随后可以消化 M-ILF 和 Peyer 斑块毛囊以进行下游细胞和分子表征。该技术需要 4-10 小时,对于对肠道免疫发育和健康和疾病功能感兴趣的研究人员很有用。

更新日期:2021-02-22
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