Creation and implantation of acellular rat renal ECM-based scaffolds

Abstract

(2015) Creation and implantation of acellular rat renal ECM-based scaffolds, Organogenesis, 11:2, 58-74, DOI: 10.1080/15476278.2015 ABSTRACT. Kidney transplantation is the only potentially curative treatment for patient facing endstage renal disease, and it is now routinely used. Its use is mainly limited by the supply of transplantable donor organs, which far exceeds the demand. Regenerative medicine and tissue engineering offer promising means for overcoming this shortage. In the present study, we developed and validated a protocol for producing acellular rat renal scaffolds.Left kidneys were removed from 26 male Lewis rats (weights: 250-350 g) and decellularized by means of aortic anterograde perfusion with ionic and anionic detergents (Triton X-100 1% and SDS 1%, respectively). 19 scaffolds thus obtained (and contralateral native kidneys as controls) were deeply characterized in order to evaluate the decellularization quality, the preservation of Ó Andrea Peloso, Jacopo Ferrario, Benedetta Maiga, Ilaria Benzoni, Carolina Bianco, Antonio Citro, Manuela Currao, Alessandro Malara, Annalisa Gaspari, Alessandra Balduini, Massimo Abelli, Lorenzo Piemonti, Paolo Dionigi, Giuseppe Orlando, and Marcello Maestri *Correspondence to: Andrea Peloso; Email: andreapeloso@hotmail.it Received August 28, 2014; Revised April 3, 2015; Accepted July 9, 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. 58 Organogenesis, 11:58-74, 2015 Published with license by Taylor & Francis Group, LLC ISSN: 1547-6278 print / 1555-8592 online DOI: 10.1080/15476278.2015 extracellular matrix components and resultant micro-angioarchitecture structure. The other 7 were transplanted into 7 recipient rats that had undergone unilateral nephrectomy. Recipients were sacrificed on post-transplantation day 7 and the scaffolds subjected to histologic studies.The dual-detergent protocol showed, with only 5 h of perfusion per organ, to obtain thoroughly decellularized renal scaffolds consisting almost exclusively of extracellular matrix. Finally the macro-and the microarchitecture of the renal parenchyma were well preserved, and the grafts were implanted with ease. Seven days after transplant, the scaffolds were morphologically intact although all vascular structures were obstructed with thrombi. Production and implantation of acellular rat renal scaffolds is a suitable platform for further studies on regenerative medicine and tissue engineering.

Biocompatibility and hemocompatibility of efficiently decellularized whole porcine kidney for tissue engineering

“…Solid organs including the heart, liver, and kidney are still challenging because of their complex architecture and their need for vasculature . Although many studies have been focused on decellularization of kidney in rodents; however, few studies described the decellularization, biocompatibility evaluation, and bioactivity of clinically large scaled porcine kidneys. Thus, in this study, we present an in‐depth analysis of decellularized porcine kidneys after optimizing a simple protocol for their decellularization and sterilization.…”

Regenerative strategies for kidney engineering

“…Also Peloso et al . observed the same problem of thrombus formation upon implantation of decellularized kidney scaffolds in a rat model. Remarkably, in a study led by Ott , whole rat acellular kidneys were obtained and recellularized with rat neonatal kidney cells and human umbilical vein endothelial cells, and further implanted orthotopically leading to some restoration of renal filtration and reabsorption.…”

“…Decellularization of whole organs has opened new perspectives on tissue engineering [78] and several laboratories have reported on the generation of renal scaffolds from rodent [79][80][81][82][83][84][85], pig [86][87][88], rhesus monkey [89] and human kidneys [5,90,91] (Table 2). A variety of decellularization protocols have been described (and reviewed elsewhere [92][93][94]) which generally involve the antegrade perfusion of detergents, enzymes or other cell-lysing solutions through the organ vasculature to remove the cellular components while preserving the 3D architecture and biochemical composition of the native ECM.…”

Bioengineering the Pancreas: Cell-on-Scaffold Technology

“…Missing ECM parts, which are not protected by vascular cells, will directly expose ECM to the blood flow, resulting in an almost instant activation of the coagulative cascade and in the formation of blood clots. This situation will bring to a subsequently blood stoppage directed to the entire portion downstream the clot, with final seeded cells death [119].…”