Kidney Week

Abstract: TH-PO0318

Global Transcriptomic Responses in Precision-Cut Kidney Slice Cultures

Session Information

• Glomerular Diseases: Omics
  October 22, 2026 | Location: Exhibit Hall A, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Glomerular Diseases

• 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

• Akilesh, Shreeram, University of Washington, Seattle, Washington, United States

Shreeram Akilesh, MD, PhD

• Li, Xianwu, University of Washington, Seattle, Washington, United States

Xianwu Li, PhD

• Beirne, Emily, University of Washington, Seattle, Washington, United States

Emily Beirne

• Mills, Bre Alexis, University of Washington, Seattle, Washington, United States

Bre Alexis Mills

• Smith, Kelly D., University of Washington, Seattle, Washington, United States

Kelly D. Smith, MD, PhD

Background

Short term human kidney slice cultures represent a tractable experimental system for studying kidney disease mechanisms. However, from the literature, the global gene expression changes that occur during ex vivo culture have not been described.

Methods

250µm vibratome slices were generated from human nephrectomy specimens and cultured in transwell inserts. Slices were cultured for various timepoints in traditional serum containing medium (medium A) or defined-component media in 95% O₂ (medium B/O₂). Slice viability was assessed using the LDH release assay. Whole transcriptome spatial gene expression profiling was performed using the GeoMx digital spatial profiler. Immunohistochemistry for NPHS1 and SLC6A19 was performed to assess protein expression.

Results

Kidney slices cultured in traditional medium in normoxia exhibited hypoxic transcriptional changes and reduced viability over 72 hrs. By contrast, kidney slices cultured in a chemically defined component medium and 95% oxygen for 72 hrs exhibited ~80% viability, though different cell stress responses were evident by transcriptional profiling. NPHS1 and SLC6A19 proteins (figures), which are sensitive markers of podocyte and proximal tubular function respectively, were both retained at 72 hrs.

Conclusion

We describe an optimized precision cut kidney slice culture protocol utilizing serum-free, chemically defined medium with increased viability and retention of markers of podocyte and proximal tubular function for up to 72 hrs ex vivo. This system will be invaluable to study kidney disease and drug mechanisms in a fully human system.