Optimized protocol for the multiomics processing of cryopreserved human kidney tissue.

Area of Science:

• Nephrology
• Translational Medicine
• Biobanking

Background:

• Clinical kidney biopsies are crucial for research but require optimized biobanking protocols.
• Current methods often necessitate liquid nitrogen, hindering integration into daily clinical workflows.
• Multi-omic analyses demand tissue preservation methods compatible with various downstream applications.

Purpose of the Study:

• To evaluate preservation media (RNAlater, CellCover) for kidney biopsy biobanking.
• To assess compatibility with multi-omic techniques including single-nucleus RNA sequencing, proteomics, and metabolomics.
• To establish protocols for integrating tissue biobanking into routine clinical practice for precision nephrology.

Main Methods:

• Modified single-nucleus dissociation protocol for snap-frozen and preserved kidney tissue.
• Single-nucleus RNA sequencing using Chromium 10X Genomics on porcine kidney tissue.
• Assessment of preservation media for proteomics, metabolomics, and histopathology (including immunofluorescence).

Main Results:

• RNAlater effectively preserves kidney tissue for single-nucleus RNA sequencing, proteomics, and histopathology.
• Transcriptomic profiles showed minimal variation across storage conditions.
• Metabolomic analysis was only feasible with snap-frozen tissue; other analyses were compatible with RNAlater.

Conclusions:

• RNAlater is a suitable preservation medium for kidney biopsies in clinical settings, enabling multi-omic analyses.
• This facilitates the creation of well-characterized biobanks for kidney disease research.
• The developed protocol supports advancements in precision medicine for kidney diseases.