Processing and cryopreservation of human ureter tissues for single-cell and spatial transcriptomics assays

Area of Science:

• Urology
• Molecular Biology
• Genomics

Background:

• Human ureter tissue characterization is crucial for understanding developmental and regenerative pathways.
• Single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics offer powerful tools for detailed cellular analysis.
• Optimized protocols are needed for reliable tissue processing for these advanced techniques.

Purpose of the Study:

• To describe optimized protocols for generating single-cell suspensions from human ureter tissue for scRNA-seq.
• To describe optimized protocols for cryopreserving human ureter tissue for 10x Genomics Visium spatial gene expression analysis.
• To facilitate comprehensive cellular and spatial mapping of the human ureter.

Main Methods:

• Single-cell RNA sequencing (scRNA-seq) protocol optimization for post-cystectomy ureter tissue.
• Cryopreservation and thawing protocol development for single-cell suspensions.
• 10x Genomics Visium spatial gene expression platform protocol optimization for cryopreserved ureter tissue.

Main Results:

• Established a robust protocol for generating high-quality single-cell suspensions from human ureter tissue.
• Developed an effective cryopreservation method for human ureter tissue suitable for spatial transcriptomics.
• These optimized protocols enable detailed cellular and spatial analyses of ureter tissue.

Conclusions:

• The described optimized protocols support advanced single-cell and spatial transcriptomic analyses of human ureter.
• These methods provide a foundation for creating a detailed cellular atlas of the human ureter.
• This work facilitates research into ureter development, regeneration, and disease.