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Isolation of myeloid cells from mouse brain tumors for single-cell RNA-seq analysis

ABSTRACT: Summary Current single-cell RNA sequencing (scRNA-seq) protocols are limited by the number of cells that can be simultaneously sequenced, restricting the ability to resolve heterogeneity of rare cell types. We describe here a protocol for rapid isolation of myeloid cells from tumor-harboring mouse cerebellum without cell sorting to minimize cell damage for scRNA-seq. This protocol includes the procedures for further enrichment of myeloid cells using CD11b+ magnetic beads, followed by the generation of scRNA library and sequencing analysis. For complete details on the use and execution of this protocol, please refer to Dang et al. (2021). Graphical abstract Highlights • Isolation of myeloid cells from tumor-bearing mouse cerebellum• Single-cell RNA sequencing (scRNA-seq) of murine cerebellar myeloid cells• Identification of the major myeloid subsets using scRNA-seq data Current single-cell RNA sequencing (scRNA-seq) protocols are limited by the number of cells that can be simultaneously sequenced, restricting the ability to resolve heterogeneity of rare cell types. We describe here a protocol for rapid isolation of myeloid cells from tumor-harboring mouse cerebellum without cell sorting to minimize cell damage for scRNA-Seq. This protocol includes the procedures for further enrichment of myeloid cells using CD11b+ magnetic beads, followed by the generation of scRNA library and sequencing analysis.

SUBMITTER: Dang M

PROVIDER: S-EPMC8605103 | biostudies-literature |

REPOSITORIES: biostudies-literature

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