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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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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) |
format | Online Article Text |
id | pubmed-10652208 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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