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Abstract: FR-PO399

Renal Tubular Epithelial Cells From Urinary Samples of Renal Transplanted Recipients: Phenotypic Characterization, Stemness, and Immunomodulatory Properties

Session Information

Category: Development‚ Stem Cells‚ and Regenerative Medicine

  • 500 Development‚ Stem Cells‚ and Regenerative Medicine

Authors

  • Pizzuti, Valeria, Nephrology, Dialysis and Renal Transplant Unit, IRCCS-Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy
  • Donadei, Chiara, Nephrology, Dialysis and Renal Transplant Unit, IRCCS-Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy
  • Gessaroli, Elisa, Nephrology, Dialysis and Renal Transplant Unit, IRCCS-Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy
  • Comai, Giorgia, Nephrology, Dialysis and Renal Transplant Unit, IRCCS-Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy
  • La Manna, Gaetano, Nephrology, Dialysis and Renal Transplant Unit, IRCCS-Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy
Background

The isolation of cells from urine samples represents a non-invasive approach to expand and characterize different cell populations. Renal Tubular Epithelial Cells (RTEC) are released into urines, and their amount can increase in the presence of kidney damage and transplant rejection.The aim of the present study was the isolation and phenotype characterization of RTEC in urinary samples from kidney transplant recipients, focusing on their stemness and immunomodulatory potential.

Methods

Urine derived RTEC, were then analyzed by cytometry for Cytokeratin, EpCAM and CD13 to confirm their epithelial features. The expression of CD90, CD73, CD44, CD105, CD24 and CD133 markers was also assayed. For the evaluation of their immunomodulatory properties, RTEC were cultured with PBMCs derived from healthy donors and the proliferation of CD4 and CD8 was analyzed with CFSE. After co-culture, T-cell subsets were quantified through flow cytometry for Th17( CD4+, IL17+), Th1 (CD4+ IFN-γ+) and Treg (CD4+CD25+, FoxP3+).

Results

The analysis of RTEC markers confirmed their epithelial features, with a high percentage of Cytokeratin positive cells . The expression of proximal and distal RTEC marker, CD13 and EpCam was observed (Fig.A,B). Most of the cells were positive for CD90, CD73, CD44, CD105, CD24 and CD133, which are typically associated with stem or progenitor populations. The co-culture with PBMCs activated with anti-CD3/CD28 showed a suppression in CD4+ and CD8+ proliferation (fig.C). Moreover, there was a reduction in Th17 and Th1 populations, while Treg subset remained stable (Fig.D).

Conclusion

The RTEC retain their epithelial characteristics after isolation and in vitro expansion. The expression of markers associated with stemness suggests the presence of not terminally differentiated subpopulations. The ability of RTEC to modulate the proliferation of immune cells could play an essential role in immune-mediated disorders.

Funding

  • Government Support – Non-U.S.