Regeneration of kidney vasculature with human kidney-derived endothelial cells in decellularized rat and human kidneys

---

D.G. Leuning, A. de Graaf, C.W. van den Berg, E. Lievers, L. Wiersma, H. de Boer, C. Avramut, B. van den Berg, C. van Kooten, M.E.J. Reinders, M. Takasoto

Chair(s): Prof. dr. C.C. Baan & dr. M.B. Rookmaaker

Wednesday 8 march 2017

17:42 - 17:54h at Hendrik Marsmanzaal

Categories: Parallel - Basaal

Parallel session: Parallelsessie V – Basaal 1 - Analyse niet-hematopoietische cellen in orgaantransplantatie

---

As there is a shortage of donor organs, there is an urgent need for new alternatives. One future alternative could be a bioengineered kidney by recellularization of a kidney scaffold with patient-derived cells. In order to achieve a functional bioengineered kidney, the vasculature should be intact. Here we study regeneration of kidney vasculature with human kidney-derived endothelial cells in both rat- and human kidney scaffolds.  

Kidney scaffolds were obtained by decellularizing rat- and human transplant grade kidneys (n=3) with 1% SDS and 0.1% TritonX-100. Preservation of structure, integrity and glycosaminoglycan (GAG) landscape of the scaffolds was analyzed with immunofluorescence for collagen type IV, fibronectin, vitronectin, laminin and heparan sulfate proteoglycans. Scaffolds were preloaded with resp. 0, 10 and 100 ng/ml vascular endothelial growth factor (VEGF).  Rat whole organ and slices of human decellularized kidney were recellularized with human primary glomerular-derived microvascular endothelial cells (hgMVEC) or human induced pluripotent stem cell-derived endothelial cells (iPS-EC). Rat kidney scaffolds were recellularized via both the renal artery and renal vein and cultured for 7 days in a custom-made organ chamber. Cell survival, adherence and coverage were analyzed by CD31 immunofluorescence and confocal microscopy.   

To conclude, human and rat kidneys showed a preservation of structure, integrity and GAG landscape after decellularization. Site specific binding of the growth factors VEGF and basic fibroblast growth factor (bFGF) was observed on these scaffolds. hgMVEC cell adherence of 20% coverage was observed in the absence of VEGF, while preloading the kidney scaffold slices with 100 ng/mL VEGF increased cell adherence to 55% coverage (P<0.001).  iPS-EC showed a similar capacity to adhere to human kidney scaffold slices. Rat decellularized kidneys showed intact vascular integrity as shown by fluorescent bead perfusion via resp. the renal artery, renal vein and urether. We show that recellularization of rat kidney scaffolds with hgMVEC via both the artery and the vein gave the highest coverage of endothelial cells in both the glomeruli and the peritubular vessels without leakage towards the tubuli.

Here we show an extensive characterization of both rat- and human kidney scaffolds for the GAG landscape and established an increase in human kidney-derived endothelial cell adherence after loading of the scaffold with VEGF. Moreover, we show a novel recellularization method of rat kidney scaffolds in an organ chamber where long term organ-culture could be achieved. These results are a promising step towards a bioengineered kidney.