Protocol for isolating immune cells from human gastric muscularis propria for single-cell analysis
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<sup>+</sup> 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.<sup>1</sup>.
Authors
- Gianluca Cipriani (ORCID: https://orcid.org/0000-0003-3267-6518)
- Shefaa AlAsfoor (ORCID: https://orcid.org/0000-0002-2179-3682)
- Gianrico Farrugia
- Madhusudan Grover (ORCID: https://orcid.org/0000-0001-5092-0831)
- Fariborz Rakhshan
- Surendra Dasari (ORCID: https://orcid.org/0000-0002-7972-3556)
- Linda C. Hsi
- Erik Jessen (ORCID: https://orcid.org/0000-0001-9851-6897)
- Kari J. Carothers
- Vernadette Simon
- Jack A. Patraw
- Julie S. Lau
- Samantha J McDonough
- Cheryl E. Bernard
Institutions
- Mayo Clinic (US)
- Mayo Clinic in Arizona (US)
- Mayo Clinic in Florida (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-08-11
- DOI
- https://doi.org/10.1016/j.xpro.2024.103258
- Citations
- 1
- Primary Topic
- Congenital gastrointestinal and neural anomalies
- Type
- article
- Field-Weighted Citation Impact
- 0.57
Funders
- Mayo Clinic
- Alexion Pharmaceuticals
- National Institute of Diabetes and Digestive and Kidney Diseases