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Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing

Single-cell isolation techniques allow the investigation of physical and functional relationships between individual cells within a complex cell population. Here, we present a protocol for single-cell isolation from full-thickness intestinal tissue resections. We describe steps for pre-processing sp...

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Autores principales: West, Gail A., Zhao, Shuai, Nguyen, Quang Tam, Christensen, Stephen M., Gordon, Ilyssa O., Holubar, Stefan D., Kravarik, Kellie M., Fiocchi, Claudio, Mukherjee, Pranab K., Rieder, Florian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10652208/
https://www.ncbi.nlm.nih.gov/pubmed/37925636
http://dx.doi.org/10.1016/j.xpro.2023.102686
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author West, Gail A.
Zhao, Shuai
Nguyen, Quang Tam
Christensen, Stephen M.
Gordon, Ilyssa O.
Holubar, Stefan D.
Kravarik, Kellie M.
Fiocchi, Claudio
Mukherjee, Pranab K.
Rieder, Florian
author_facet West, Gail A.
Zhao, Shuai
Nguyen, Quang Tam
Christensen, Stephen M.
Gordon, Ilyssa O.
Holubar, Stefan D.
Kravarik, Kellie M.
Fiocchi, Claudio
Mukherjee, Pranab K.
Rieder, Florian
author_sort West, Gail A.
collection PubMed
description Single-cell isolation techniques allow the investigation of physical and functional relationships between individual cells within a complex cell population. Here, we present a protocol for single-cell isolation from full-thickness intestinal tissue resections. We describe steps for pre-processing specimens, isolation of lamina propria and muscular layers, and red blood cell lysis. We then detail fixation of isolated cells and assessment of cell quality. The resulting cell suspension can be subjected to RNA sequencing on the 10× Chromium platform. For complete details on the use and execution of this protocol, please refer to Mukherjee et al.(1)
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spelling pubmed-106522082023-11-04 Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing West, Gail A. Zhao, Shuai Nguyen, Quang Tam Christensen, Stephen M. Gordon, Ilyssa O. Holubar, Stefan D. Kravarik, Kellie M. Fiocchi, Claudio Mukherjee, Pranab K. Rieder, Florian STAR Protoc Protocol Single-cell isolation techniques allow the investigation of physical and functional relationships between individual cells within a complex cell population. Here, we present a protocol for single-cell isolation from full-thickness intestinal tissue resections. We describe steps for pre-processing specimens, isolation of lamina propria and muscular layers, and red blood cell lysis. We then detail fixation of isolated cells and assessment of cell quality. The resulting cell suspension can be subjected to RNA sequencing on the 10× Chromium platform. For complete details on the use and execution of this protocol, please refer to Mukherjee et al.(1) Elsevier 2023-11-04 /pmc/articles/PMC10652208/ /pubmed/37925636 http://dx.doi.org/10.1016/j.xpro.2023.102686 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
West, Gail A.
Zhao, Shuai
Nguyen, Quang Tam
Christensen, Stephen M.
Gordon, Ilyssa O.
Holubar, Stefan D.
Kravarik, Kellie M.
Fiocchi, Claudio
Mukherjee, Pranab K.
Rieder, Florian
Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing
title Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing
title_full Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing
title_fullStr Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing
title_full_unstemmed Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing
title_short Single-cell isolation from full-thickness human intestinal tissue resections for single-cell RNA sequencing
title_sort single-cell isolation from full-thickness human intestinal tissue resections for single-cell rna sequencing
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10652208/
https://www.ncbi.nlm.nih.gov/pubmed/37925636
http://dx.doi.org/10.1016/j.xpro.2023.102686
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