Publication: Development of decellularization protocols for female cat reproductive organs
Authors
Sanguansook, P ; Martínez-López, Cristina ; López-Orozco, Marina ; Chatdarong, K ; García-Vázquez, FA ; Martínez Cáceres, Carlos Manuel ; Izquierdo Rico, María José
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Publisher
Elsevier
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DOI
https://doi.org/10.1016/j.rvsc.2024.105257
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info:eu-repo/semantics/article
Description
© 2024 Authors
This document is the published version of a published work that appeared in final form in Research in Veterinary Science
This document is made available under the CC-BY 4.0 license http://creativecommons.org/licenses/by/4.0
To access the final edited and published work see:
https://doi.org/10.1016/j.rvsc.2024.105257
Abstract
Decellularization is an innovative method to create natural scaffolds by removing all cellular materials while
preserving the composition and three-dimensional ultrastructure of the extracellular matrix (ECM). The obtention
of decellularized reproductive organs in cats might facilitate the development of assisted reproductive
techniques not only in this species but also in other felids. The aim was to compare the efficiency of three
decellularization protocols on reproductive organs (ovary, oviduct, and uterine horn) in domestic cats. The
decellularization protocol involved 0.1% sodium dodecyl sulfate and 1%Triton X-100. Protocol 1 (P1) entailed 2-
cycles of decellularization using these detergents. Protocol 2 (P2) was like P1 but included 3-cycles. Protocol 3
(P3) was similar to P2, with the addition of deoxyribonuclease incubation. Reproductive organs from nine cats
were separated into two sides. One side served as the control (non-decellularized organ) while the contralateral
side was the treated group (decellularized organ). The treated organs were subdivided into 3 groups (n = 3 per
group) for each protocol. Both control and treated samples were analyzed for DNA content, histology (nuclear
and ECM (collagen, elastin, and glycosaminoglycans (GAGs)) density), ultrastructure by electron microscopy,
and cytotoxicity. The results of the study showed that P3 was the only protocol that displayed no nucleus residue
and significantly reduced DNA content in decellularized samples (in all the studied organs) compared to the
control (P < 0.05). The ECM content in the ovaries remained similar across all protocols compared with controls
(P > 0.05). However, elastic fibers and GAGs decreased in decellularized oviducts (P < 0.05), while collagen
levels remained unchanged (P > 0.05). Regarding the uterus, the ECM content decreased in decellularized
uterine horns from P3 (P < 0.05). Electron microscopy revealed that the microarchitecture of the decellularized
samples was maintained compared to controls. The decellularized tissues, upon being washed for 24 h, showed
cytocompatibility following co-incubation with sperm. In conclusion, when comparing different decellularization
methods, P3 proved to be the most efficient in removing nuclear material from reproductive organs compared to
P1 and P2. P3 demonstrated its success in decellularizing ovarian samples by significantly decreasing DNA
content while maintaining ECM components and tissue microarchitecture. However, P3 was less effective in
maintaining ECM contents in decellularized oviducts and uterine horns.
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Citation
Research in Veterinary Science 173 (2024) 105257
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