Isolation of macrophages from mouse skin wounds for single-cell RNA sequencing

STAR Protocols

Susanne Reinhardt¹  Andreas Dahl²  Juliana G. Roscito²  Sebastian Willenborg³  Sabine A. Eming³  Axel Roers⁴  Alexander Gerbaulet⁴ 

[1] Center for Molecular Medicine Cologne (CMMC), University of Cologne, 50931 Cologne, Germany;DRESDEN-concept Genome Center (DcGC), Center for Molecular and Cellular Bioengineering, TU Dresden, 01307 Dresden, Germany;Department of Dermatology, University of Cologne, 50937 Cologne, Germany;Institute for Immunology, Medical Faculty, TU Dresden, 01307 Dresden, Germany;

关键词: Cell Biology;    Cell isolation;    Single Cell;    Flow Cytometry/Mass Cytometry;    Sequencing;    RNAseq;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: Understanding macrophage heterogeneity in tissue repair is a major challenge. Here, we describe a protocol that combines isolation of immune cells from skin wounds with subsequent flow-cytometry-based sorting of wound macrophages and single-cell RNA sequencing. We use a modified version of the original Smart-seq2 protocol to increase speed and accuracy. This protocol is useful for analyzing the pronounced heterogeneity of activation phenotypes in wound macrophages and might be adapted to other experimental models of skin inflammation.For complete details on the use and execution of this protocol, please refer to Willenborg et al. (2021).

【 授权许可】

Unknown