Human Endothelial Cell Seeding in Partially Decellularized Kidneys

Abstract

The excessive demand for organ transplants has promoted the development of strategies that increase the supply of immune compatible organs, such as xenotransplantation of genetically modified pig organs and the generation of bioartificial organs. We describe a method for the partial replacement of rat endothelial cells for human endothelial cells in a rat’s kidney, obtaining as a final result a rat-human bioartificial kidney. First, in order to maintain parenchymal epithelial cells and selectively eliminate rat endothelial cells, three methods were evaluated in which different solutions were perfused through the renal artery: 0.1% sodium dodecyl sulfate (SDS), 0.01% SDS, and hyperosmolar solutions of sucrose. Then, partially decellularized kidneys were recellularized with human endothelial cells and finally transplanted in an anesthetized rat. The solution of 0.1% SDS achieved the highest vascular decellularization but with high degree of damage in the parenchyma side. On the contrary, 0.01% SDS and hyperosmolar solutions achieved a partial degree of endothelial decellularization. TUNEL assays reveal that hyperosmolar solutions maintained a better epithelial cell viability contrasting with 0.01% SDS. Partially decellularized kidneys were then recellularized with human endothelial cells. Histological analysis showed endothelial cells attached in almost all the vascular bed. Recellularized kidney was transplanted in an anesthetized rat. After surgery, recellularized kidney achieved complete perfusion, and urine was produced for at least 90 min posttransplant. Histological analysis showed endothelial cells attached in almost all the vascular bed. Therefore, endothelial decellularization of grafts and recellularization with human endothelial cells derived from transplant recipients can be a feasible method with the aim to reduce the damage of the grafts.

Bioengineering of vascularized porcine flaps using perfusion-recellularization

“…This is in alignment with observed venous return in decellularized omentum and TFL previously published by our group. Haeublein et al similarly used Evan’s blue dye to quantify vascular permeability following decellularization of the rat kidney at various SDS concentrations 36 . Although they noted an increase in vascular permeability following decellularization at both high and low SDS concentrations, the low-concentration SDS had lower loss of permeability.…”

Angiogenesis and Re-endothelialization in decellularized scaffolds: Recent advances and current challenges in tissue engineering

“…As a result, the perfused tissue will have a leaky structure in the absence of an efficient venous and lymphatic system ( Ren et al, 2015 ). Studies of the continuity and integrity of the vascular bed in acellular rat lungs and kidneys have revealed a degree of vascular permeability, media leakage, and a reduction in hydrostatic pressure along the vessels ( Stabler et al, 2016 ; Haeublein et al, 2022 ). This leads to media leakage into the parenchyma, reducing the efficiency of cell delivery and diminishing the endothelialization rate ( Stabler et al, 2016 ).…”

“…Multiple strategies and processes have been adopted for this goal and to respond to the challenges described above, each of which separately plays a vital role in the endothelialization process ( Seiffert et al, 2021 ; Daryabari et al, 2022 ; Haeublein et al, 2022 ; Kim et al, 2022 ). Decellularization and its protocols are among the first influential factors in the endothelialization of acellular structures.…”

Research progress of biomaterials and innovative technologies in urinary tissue engineering

“…After 9 months, complete regeneration of the bladder wall was achieved by implanting the decellularized ureters, and the boundary between the host bladder and the decellularized ureter was no longer easily distinguished. The decellularized urethral grafts in this study have broad potential as biocompatible scaffolds for cell ingrowth and morphological formation of bladder tissues ( Singh et al, 2018 ; Haeublein et al, 2022 ).…”