Comparative evaluation of ACetic - MEthanol high salt dissociation approach for single-cell transcriptomics of frozen human tissues.

Summary

The authors optimize an acetic acid–methanol dissociation with high-salt washes (ACME HS) to recover intact cells from fresh-frozen human endocrine tissues for scRNA-seq, preserving morphology and RNA integrity. Across 41 samples, ACME HS compared favorably with enzymatic dissociation and nuclei isolation for cell-type preservation and standard QC metrics.

Key Findings

• ACME HS (acetic acid–methanol with high-salt wash) preserves cell morphology and RNA integrity from fresh-frozen endocrine tissues.
• Across 41 samples, ACME HS maintains major cell types and gene expression profiles with strong QC metrics compared to enzymatic dissociation and nuclei isolation.
• High-salt buffer during rehydration reduces RNase reactivation, stabilizing RNA and minimizing transcriptomic artifacts.

Clinical Implications

While not directly clinical, ACME HS can accelerate translational discovery by enabling scalable, less biased single-cell atlases of human endocrine organs (e.g., thyroid, pancreas), informing disease mechanisms and targets.

Limitations

• Primarily a methods paper without clinical outcomes; generalizability beyond tested tissues requires further validation.
• Performance across diverse fixation durations and storage conditions was not exhaustively characterized.

Future Directions

Benchmark ACME HS across endocrine disease tissues (e.g., thyroid cancer, islet autoimmunity), integrate with multi-omics, and standardize for biobank protocols.