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Identification, isolation and analysis of human gut-associated lymphoid tissues

Jørgensen, Peter B. ; Fenton, Thomas M. ; Mörbe, Urs M. LU ; Riis, Lene B. ; Jakobsen, Henrik L. ; Nielsen, Ole H. and Agace, William W. LU (2021) In Nature Protocols 16(4). p.2051-2067
Abstract

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... (More)

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.

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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Nature Protocols
volume
16
issue
4
pages
17 pages
publisher
Nature Publishing Group
external identifiers
  • scopus:85101262285
  • pmid:33619391
ISSN
1754-2189
DOI
10.1038/s41596-020-00482-1
language
English
LU publication?
yes
id
354ba10d-44bc-476a-8702-ac7f0dbd95b8
date added to LUP
2021-03-10 14:40:21
date last changed
2024-09-05 16:45:54
@article{354ba10d-44bc-476a-8702-ac7f0dbd95b8,
  abstract     = {{<p>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.</p>}},
  author       = {{Jørgensen, Peter B. and Fenton, Thomas M. and Mörbe, Urs M. and Riis, Lene B. and Jakobsen, Henrik L. and Nielsen, Ole H. and Agace, William W.}},
  issn         = {{1754-2189}},
  language     = {{eng}},
  month        = {{04}},
  number       = {{4}},
  pages        = {{2051--2067}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Nature Protocols}},
  title        = {{Identification, isolation and analysis of human gut-associated lymphoid tissues}},
  url          = {{http://dx.doi.org/10.1038/s41596-020-00482-1}},
  doi          = {{10.1038/s41596-020-00482-1}},
  volume       = {{16}},
  year         = {{2021}},
}