Protocol to dissociate, process, and analyze the human lung tissue using single-cell RNA-seq

Abstract

Summary: We report a protocol for obtaining high-quality single-cell transcriptomics data from human lung biospecimens acquired from core needle biopsies, fine-needle aspirates, surgical resection, and pleural effusions. The protocol relies upon the brief mechanical and enzymatic disruption of tissue, enrichment of live cells by fluorescence-activated cell sorting (FACS), and droplet-based single-cell RNA sequencing (scRNA-seq). The protocol also details a procedure for analyzing the scRNA-seq data.For complete details on the use and execution of this protocol, please refer to Chan et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cancer
• Cell isolation
• Flow cytometry/Mass cytometry
• Molecular biology
• RNAseq
• Sequence analysis