Abstract: SA-PO0030
Establishment and Characterization of a Patient-Derived Proximal Tubular Cell Culture Model from Human Kidney Biopsies
Session Information
- Development, Stem Cells, and Regenerative Medicine
October 24, 2026 | Location: Exhibit Hall A, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 700 Development, Stem Cells, and Regenerative Medicine
Authors
- Mohamed, Mai Abdelhalim, Medizinische Hochschule Hannover, Hanover, NDS, Germany
- Tsamo Tetou, Arnold, Medizinische Hochschule Hannover, Hanover, NDS, Germany
- Günzel, Dorothee, Charite - Universitatsmedizin Berlin, Berlin, BE, Germany
- Lovric, Svjetlana, Medizinische Hochschule Hannover, Hanover, NDS, Germany
- Hiss, Marcus, Medizinische Hochschule Hannover, Hanover, NDS, Germany
- López-Cayuqueo, Karen I., Medizinische Hochschule Hannover, Hanover, NDS, Germany
- Schmidt-Ott, Kai M., Medizinische Hochschule Hannover, Hanover, NDS, Germany
Background
Patient-derived primary proximal tubular cell culture models are constrained by the scant tissue yields of routine diagnostic kidney biopsies. This study aimed to establish a reproducible protocol for generating primary human proximal tubular cell cultures from ultra-small biopsy specimens and to define their cellular identity.
Methods
Fresh ultrasound-guided human kidney biopsies from chronic kidney disease patients were collected and microscopically inspected to identify the cortical region. Approximately 0.5 cm of cortical tissue underwent collagenase-based digestion. Post enzymatic digestion, the resultant cell pellet was filtered and seeded as a heterogeneous mixture of single cells and tubular fragments. Following a 7-day expansion in renal epithelial growth medium, cultures underwent multi-modal phenotypic characterization using morphological assessment, protein expression analysis, and transcript profiling by single-cell RNA sequencing.
Results
Minimal biopsy material reproducibly generated adherent epithelial monolayers within 7 days. Cultured cells displayed typical cobblestone morphology and active proliferation. Immunofluorescence demonstrated predominant Megalin expression with marked BrdU incorporation, indicating proximal tubular identity and active proliferative capacity. Gene expression analysis revealed enrichment of the proximal tubular marker LRP2, whereas expression levels of distal nephron markers AQP2, SLC12A1 (NKCC2), and SLC12A3 (NCC) remained comparatively low in primary cultures. Single-cell RNA sequencing identified predominant proximal tubular populations (70%) with minor endothelial and stromal cell subsets.
Conclusion
We developed a robust methodology for deriving primary human proximal tubular cell cultures from ultra-small diagnostic kidney biopsies while preserving key tubular phenotypic features. By demonstrating the feasibility of utilizing minimal samples, this clinically relevant patient-derived model may provide a translational platform for disease modelling, therapeutic screening, and mechanistic studies of tubular injury.
Funding
- Government Support – Non-U.S.