PKHhigh/CD133+/CD24- Renal Stem-Like Cells Isolated from Human Nephrospheres Exhibit In Vitro Multipotency.

Silvia Bombelli,Chiara Meregalli,Antonino Bruno,Adriana Albini,Davide P Bernasconi,Paolo Mazzola,Nicola Zucchini,Chiara Grasselli,Cristina Bianchi,Maddalena M Bolognesi,Giorgio Cattoretti,Sofia De Marco,Marco Grasso,Stefano Eriani,Roberto A Perego,Barbara Torsello

doi:10.3390/cells9081805

Abstract

The mechanism upon which human kidneys undergo regeneration is debated, though different lineage-tracing mouse models have tried to explain the cellular types and the mechanisms involved. Different sources of human renal progenitors have been proposed, but it is difficult to argue whether these populations have the same capacities that have been described in mice. Using the nephrosphere (NS) model, we isolated the quiescent population of adult human renal stem-like PKHhigh/CD133+/CD24− cells (RSC). The aim of this study was to deepen the RSC in vitro multipotency capacity. RSC, not expressing endothelial markers, generated secondary nephrospheres containing CD31+/vWf+ cells and cytokeratin positive cells, indicating the coexistence of endothelial and epithelial commitment. RSC cultured on decellularized human renal scaffolds generated endothelial structures together with the proximal and distal tubular structures. CD31+ endothelial committed progenitors sorted from nephrospheres generated spheroids with endothelial-like sprouts in Matrigel. We also demonstrated the double commitment toward endothelial and epithelial lineages of single RSC. The ability of the plastic RSC population to recapitulate the development of tubular epithelial and endothelial renal lineages makes these cells a good tool for the creation of organoids with translational relevance for studying the parenchymal and endothelial cell interactions and developing new therapeutic strategies.

Highlights

Neonephrogenesis does not occur in the adult human kidney, but the kidney retains some regenerative potential to replace the loss of cells during physiological processes, in which about 70,000 cells fromCells 2020, 9, 1805; doi:10.3390/cells9081805 www.mdpi.com/journal/cellsCells 2020, 9, 1805 different nephron segment are lost every hour and excreted with urine [1]
renal stem-like PKHhigh/CD133+/CD24− cells (RSC) (PKHhigh /CD133+/CD24− cells) were obtained by FACS sorting from NS cultured for 10 days
Subpopulation, able to generate NS, had the capacity to attach to tubular and vascular segments of renal scaffolds differentiating in epithelial- and endothelial-like lineages, respectively

Summary

Introduction

Neonephrogenesis does not occur in the adult human kidney, but the kidney retains some regenerative potential to replace the loss of cells during physiological processes, in which about 70,000 cells fromCells 2020, 9, 1805; doi:10.3390/cells9081805 www.mdpi.com/journal/cellsCells 2020, 9, 1805 different nephron segment are lost every hour and excreted with urine [1]. It has long been known that in the kidney some tubular cells can undergo hypertrophy and some hyperplasia [5]. It remained to be determined whether any differentiated tubular cell under specific conditions is capable to give rise to dedifferentiation and clonal proliferation of tubular cells or if only a specific predetermined fraction of cells (stem/progenitor cells) is endowed with this potential within the nephron [6,7]. Lazzeri and colleagues [8] identified and characterized the tubular cells that undergo the endocycle-mediated hypertrophy and the small subset of Pax2+ tubular progenitor cells that have proliferative capacity in conditional Pax8/FUCCI2aR mice

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