Dissociation of microdissected mouse brain tissue for artifact free single-cell RNA sequencing

Abstract

Summary: Single-cell RNA sequencing (scRNA-seq) provides the transcriptome of individual cells and addresses previously intractable problems including the central nervous system’s transcriptional responses during health and disease. However, dissociating brain cells is challenging and induces artificial transcriptional responses. Here, we describe an enzymatic dissociation method for mouse brain that prevents dissociation artifacts and lowers technical variations with standardized steps. We tested this protocol on microdissected brain tissue of 3-week- to 24-month-old mice and obtained high-quality scRNA-seq results.For complete details on the use and execution of this protocol, please refer to Safaiyan et al. (2021).

Keywords

• Cell Biology
• Cell isolation
• Single Cell
• Flow Cytometry/Mass Cytometry
• Sequencing
• RNA-seq