Division of Neurology, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
Division of Pediatric and Developmental Neurology, Washington University School of Medicine, St. Louis, MO 63110, USA.
STAR Protoc. 2021 Nov 17;2(4):100957. doi: 10.1016/j.xpro.2021.100957. eCollection 2021 Dec 17.
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).
目前的单细胞 RNA 测序 (scRNA-seq) 方案受到可同时测序的细胞数量的限制，限制了对稀有细胞类型异质性的解析能力。我们在这里描述了一种从携带肿瘤的小鼠小脑快速分离髓样细胞的方案，而无需进行细胞分选以最大程度减少 scRNA-seq 中的细胞损伤。该方案包括使用 CD11b+磁珠进一步富集髓样细胞的步骤，然后进行 scRNA 文库的生成和测序分析。有关此方案的使用和执行的完整详细信息，请参阅 Dang 等人（2021 年）。
