Protocol for isolating immune cells from human gastric muscularis propria for single-cell analysis.

Autor:	AlAsfoor S; Division of Gastroenterology and Hepatology, Enteric Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA. Electronic address: alasfoor.shefaa@mayo.edu., Bernard CE; Division of Gastroenterology and Hepatology, Enteric Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA., Jessen E; Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN 55905, USA., Simon VA; Center for Individualized Medicine (CIM), Mayo Clinic, Rochester, MN 55905, USA., Rakhshan F; Center for Individualized Medicine (CIM), Mayo Clinic, Rochester, MN 55905, USA., Cipriani G; Division of Gastroenterology and Hepatology, Enteric Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA., Hsi LC; Division of Gastroenterology and Hepatology, Enteric Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA., Patraw JA; Division of Gastroenterology and Hepatology, Enteric Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA., Carothers KJ; Center for Individualized Medicine (CIM), Mayo Clinic, Rochester, MN 55905, USA., Lau JS; Center for Individualized Medicine (CIM), Mayo Clinic, Rochester, MN 55905, USA., McDonough SJ; Center for Individualized Medicine (CIM), Mayo Clinic, Rochester, MN 55905, USA., Dasari S; Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN 55905, USA., Farrugia G; Division of Gastroenterology and Hepatology, Enteric Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA., Grover M; Division of Gastroenterology and Hepatology, Enteric Neuroscience Program, Mayo Clinic, Rochester, MN 55905, USA. Electronic address: grover.madhusudan@mayo.edu.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2024 Sep 20; Vol. 5 (3), pp. 103258. Date of Electronic Publication: 2024 Aug 11.
DOI:	10.1016/j.xpro.2024.103258
Abstrakt:	Understanding the diversity of gastrointestinal (GI) immune cells, especially in the muscularis propria, is crucial for understanding their role in the maintenance of enteric neurons and smooth muscle and their contribution to GI motility. Here, we present a detailed protocol for isolating single immune cells from the human gastric muscularis propria. We describe steps for tissue preservation, dissection, and dissociation of the muscularis propria. We then detail procedures for magnetic sorting of CD45 + cells and single-cell RNA sequencing (scRNA-seq) analysis. For complete details on the use and execution of this protocol, please refer to Chikkamenahalli et al. 1 . Competing Interests: Declaration of interests M.G. has received investigator-initiated grant funding from Takeda Pharmaceuticals and clinical trial funding from Alexza and Donga Pharmaceuticals. He serves as a consultant for Alfasigma, Ardelyx, and Evoke Pharmaceuticals. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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