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An optimized protocol for the isolation of rare stromal cell populations from the mouse spleen

ABSTRACT: Summary Lymphoid tissue stromal cells are important regulators of spleen homeostasis and immune responses. Here, we present an optimized protocol that describes the digestion and enrichment steps for the isolation and analysis of rare populations of stromal cells, including fibroblastic reticular cells, perivascular cells, and glial cells found in the spleen. This protocol is suitable for subsequent analysis of spleen stromal cells by flow cytometry or single-cell RNA sequencing and to analyze different disease models. For complete details on the use and execution of this protocol, please refer to Alexandre et al. (2022).1 Graphical abstract Highlights • Protocol for the isolation and analysis of mouse splenic stromal cells• Digestion method to release stromal cells• Enrichment and antibody staining of stromal cells• Details of FACS analysis of splenic stromal cell subsets Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Lymphoid tissue stromal cells are important regulators of spleen homeostasis and immune responses. Here, we present an optimized protocol that describes the digestion and enrichment steps for the isolation and analysis of rare populations of stromal cells, including fibroblastic reticular cells, perivascular cells, and glial cells found in the spleen. This protocol is suitable for subsequent analysis of spleen stromal cells by flow cytometry or single-cell RNA sequencing and to analyze different disease models.

SUBMITTER: Alexandre Y

PROVIDER: S-EPMC9768415 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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An optimized protocol for the isolation of rare stromal cell populations from the mouse spleen

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