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

Area of Science:

• Pulmonology
• Molecular Biology
• Genomics

Background:

• Single-cell transcriptomics offers deep insights into lung tissue heterogeneity.
• Existing protocols may not be optimal for diverse human lung specimen types.

Purpose of the Study:

• To establish a robust protocol for high-quality single-cell RNA sequencing (scRNA-seq) from various human lung biospecimens.
• To enable comprehensive analysis of cellular composition and function in lung diseases.

Main Methods:

• Tissue dissociation using brief mechanical and enzymatic disruption.
• Live cell enrichment via fluorescence-activated cell sorting (FACS).
• Droplet-based single-cell RNA sequencing (scRNA-seq) and data analysis pipeline.

Main Results:

• Successful generation of high-quality scRNA-seq data from core needle biopsies, fine-needle aspirates, surgical resections, and pleural effusions.
• Demonstration of the protocol's efficacy across different human lung tissue sources.
• Detailed procedure for subsequent scRNA-seq data analysis.

Conclusions:

• The developed protocol provides a reliable method for single-cell transcriptomic analysis of human lung samples.
• This facilitates deeper understanding of lung biology and pathology at the single-cell level.
• The protocol supports diverse clinical applications in respiratory research.