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Merck
CN
  • An optimized protocol for the isolation of rare stromal cell populations from the mouse spleen.

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

STAR protocols (2023-01-04)
Yannick O Alexandre, Scott N Mueller
摘要

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.

材料
Product Number
品牌
产品描述

Roche
胶原酶D, from Clostridium histolyticum
Sigma-Aldrich
脱氧核糖核酸酶 I 来源于牛胰腺, lyophilized powder, Protein ≥85 %, ≥400 Kunitz units/mg protein
Sigma-Aldrich
乙二胺四乙酸 二钠盐 二水合物, suitable for electrophoresis, Molecular Biology, 99.0-101.0% (titration)
Sigma-Aldrich
DAPI, for nucleic acid staining
Roche
分散酶®II (中性蛋白酶, II级), lyophilized, from bacterial, Roche, pkg of 5 × 1 g