Decellularization of canine kidney for three-dimensional organ regeneration

Vet World Vol.13 March-2020 Article-9

Research Article

Veterinary World, 13(3): 452-457

https://doi.org/10.14202/vetworld.2020.452-457

Kazuki Tajima^1,2, Kohei Kuroda², Yuya Otaka¹, Rie Kinoshita², Mizuki Kita¹, Toshifumi Oyamada³, and Kazutaka Kanai¹

1. Department of Small Animal Internal Medicine II, School of Veterinary Medicine, Kitasato University, Towada, Japan.
2. Department of Surgery, Keio University School of Medicine, Shinjuku, Japan.
3. Department of Veterinary Pathology, School of Veterinary Medicine, Kitasato University, Towada, Japan.

Background and Aim: Kidney regeneration is required for dogs with end-stage renal failure. Decellularization is one of the bioengineering techniques, which involves the removal of all tissue cells and cellular components and conservation of the extracellular matrix (ECM). Studies in rats have shown that decellularized kidney has regenerative potential; however, there are no reports on renal decellularization in dogs. Here, we showed the decellularization of the canine kidney.

Materials and Methods: The renal artery of the cadaveric canine kidney was cannulated and the whole kidney was frozen at –80°C. After completely thawing, it was perfused with physiological saline and sodium dodecyl sulfate (0.5%, 6 h) through the cannulated renal artery to achieve decellularization. To assess the efficiency of the decellularization protocol, histological and immunohistochemical analysis of decellularized kidney was performed.

Results: The results of hematoxylin and eosin (H and E) staining revealed that the decellularized canine kidney had no apparent cellular components. In addition, 4’,6-diamidino-2-phenylindole (DAPI) staining showed no visible nuclear components within the whole decellularized kidney. Therefore, both H and E and DAPI staining showed decellularization of the canine kidney. Our decellularization protocol also preserved the basement membrane of glomerulus, shown by periodic acid methenamine silver, periodic acid–Schiff, fibronectin, and collagen type IV stain.

Conclusion: Our decellularization protocol could eliminate cellular components and remaining native ECM structures of canine kidney. These results could promote further research into canine kidney regeneration, which may be the first small step to regenerate the canine kidney waiting for renal transplantation. Keywords: bioengineering, dog, extracellular matrix, kidney, regeneration.

Keywords: bioengineering, dog, extracellular matrix, kidney, regeneration.

How to cite this article: Tajima K, Kuroda K, Otaka Y, Kinoshita R, Kita M, Oyamada T, Kanai K (2020) Decellularization of canine kidney for three-dimensional organ regeneration, Veterinary World, 13(3): 452-457.

Received: 27-11-2019 Accepted: 04-02-2020 Published online: 12-03-2020

Corresponding author: Kazutaka Kanai E-mail: kanai@vmas.kitasato-u.ac.jp

DOI: 10.14202/vetworld.2020.452-457

Copyright: Tajima, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.