@article {20295, title = {Co-cultures of renal progenitors and endothelial cells on kidney decellularized matrices replicate the renal tubular environment in vitro}, journal = {Acta Physiologica}, year = {2020}, month = {2020-05-04 00:00:00}, publisher = {John Wiley \& Sons, Ltd}, abstract = {

Aim

Herein we propose creating a bilayer tubular kidney\ in-vitro\ model. It is hypothesized that membranes composed of decellularized porcine kidney extracellular matrix are valid substitutes of the tubular basement membrane by mimicking the physiological relevance of the\ in vivo\ environment and disease phenotypes.

Methods

Extracellular matrix was obtained from decellularized porcine kidneys. After processing by lyophilization and milling, it was dissolved in an organic solvent and blended with poly(caprolactone). Porous membranes were obtained by electrospinning and seeded with human primary renal progenitor cells to evaluate phenotypic alterations. To create a bilayer model of the\ in vivo\ tubule, the same cells were differentiated into epithelial tubular cells and co-cultured with endothelial cells in opposite sites.

Results

Our results demonstrate increasing metabolic activity, proliferation and total protein content of renal progenitors over time. We confirmed the expression of several genes encoding epithelial transport proteins and we could also detect tubular-specific proteins by immunofluorescence stainings. Functional and transport assays were performed trough the bilayer by quantifying both human serum albumin uptake and inulin leakage. Furthermore, we validated the chemical modulation of nephrotoxicity on this epithelium-endothelium model by cisplatin exposure.

Conclusion

The use of decellularized matrices in combination with primary renal cells was shown to be a valuable tool for modeling renal function and disease\ in-vitro. We successfully validated our hypothesis by replicating the physiological conditions of an\ in vitro\ tubular bilayer model. The developed system may contribute significantly for the future investigation of advanced therapies for kidney diseases.

}, keywords = {Cellular crosstalk, Decellularization, Extracellular matrix, Kidney, Renal progenitors, Tubule}, issn = {1748-1708}, doi = {10.1111/apha.13491}, url = {https://onlinelibrary.wiley.com/doi/abs/10.1111/apha.13491}, author = {Sobreiro-Almeida, R. and Melica, M. E. and Lasagni, L. and Romagnani, P. and Neves, N. M.} }