Single-cell transcriptome conservation in cryopreserved cells and tissues.

Area of Science:

• Molecular Biology
• Genomics
• Biotechnology

Background:

• Current single-cell RNA sequencing (scRNA-seq) protocols typically require fresh biological samples.
• The need for immediate processing limits the feasibility of large-scale or geographically dispersed study designs.
• Existing methods for sample preservation often compromise RNA integrity, affecting transcriptomic analysis.

Purpose of the Study:

• To develop and validate a sample preservation method for single-cell RNA sequencing.
• To enable the decoupling of sample collection from laboratory processing.
• To expand the applicability of scRNA-seq for complex and large-scale research.

Main Methods:

• A novel method for preserving single cells while maintaining RNA transcripts was developed.
• Single-cell transcriptomes were analyzed from over 1000 fresh and cryopreserved cells.
• Both 3'-end and full-length RNA preparation methods were employed for sequencing.

Main Results:

• The developed preservation method successfully maintained RNA integrity in single cells.
• Transcriptional profiles of preserved cells were comparable to those of fresh cells.
• No significant alterations in transcriptional profiles were observed due to the conservation process.

Conclusions:

• The described sample preservation technique is effective for single-cell transcriptomics.
• This method overcomes the limitations of fresh-sample requirements, facilitating complex study designs.
• The findings suggest a potential paradigm shift in the methodology and scope of future scRNA-seq studies.