Identification, isolation and analysis of human gut-associated lymphoid tissues
(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
- Jørgensen, Peter B. ; Fenton, Thomas M. ; Mörbe, Urs M. LU ; Riis, Lene B. ; Jakobsen, Henrik L. ; Nielsen, Ole H. and Agace, William W. LU
- organization
- publishing date
- 2021-04-01
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Nature Protocols
- volume
- 16
- issue
- 4
- pages
- 17 pages
- publisher
- Nature Publishing Group
- external identifiers
-
- pmid:33619391
- scopus:85101262285
- 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}}, }