CryoSCAPE: Scalable immune profiling using cryopreserved whole blood for multi-omic single cell and functional assays

Area of Science:

• Immunology
• Single-cell genomics
• Proteomics

Background:

• Single-cell technologies have significantly advanced understanding of immune system heterogeneity and function.
• Cryopreservation of peripheral blood mononuclear cells (PBMC) is crucial for immune cell characterization but faces clinical accessibility challenges.
• CryoSCAPE (Cryopreservation for Scalable Cellular And Proteomic Exploration) offers a scalable solution using direct whole blood cryopreservation for immune studies.

Purpose of the Study:

• To develop and validate a scalable method for single-cell immune analysis using direct whole blood cryopreservation.
• To assess the compatibility of cryopreserved whole blood with multi-omic single-cell assays and functional assays.
• To overcome limitations in clinical isolation and preservation of immune cells for research.

Main Methods:

• Direct cryopreservation of whole blood for subsequent immune cell isolation and multi-omic single-cell assays.
• Comparative analysis of immune cell proportions and molecular features between cryopreserved whole blood and density gradient isolated PBMC.
• Optimization and validation for high sample throughput using automated liquid handling and fixed single-cell RNA sequencing.

Main Results:

• Cryopreserved whole blood effectively captures immune cell proportions and molecular features comparable to density gradient isolated PBMC.
• The CryoSCAPE method was optimized and validated for high-throughput analysis using automated systems with large sample batches.
• Cryopreserved whole blood is compatible with functional assays, demonstrating its utility for clinical research.

Conclusions:

• CryoSCAPE provides a scalable, cost-effective method for high-throughput single-cell RNA sequencing and functional assays.
• The method simplifies sample handling, potentially democratizing access to single-cell immune assays.
• CryoSCAPE can enhance the understanding of immune function across diverse populations by improving research accessibility.