Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2025-0096
Animal Reproduction (AR)
ORIGINAL ARTICLE

Cryopreservation of jaguar (Panthera onca) fibroblast-like cells in straws and cryovials: a comparison of rapid and slow-freezing protocols

Sofia Regina Polizelle; Ian Figueiredo Navarezi; Maite Cardoso Coelho da Silva; Giulia Correia Vieira; Liandra Martiliano Gaeth; Larissa Schneider Brandão-Souza; Bianca Rodrigues Acacio; Pedro Nacib Jorge-Neto; Eliane Vianna da Costa e Silva; Thyara de Deco-Souza; Alexsandra Fernandes Pereira; Gediendson Ribeiro de Araujo

http://dx.doi.org/10.1590/1984-3143-AR2025-0096 Anim Reprod, vol.23, n3, e20250096, 2026
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Abstract

Somatic cells serve as essential genetic resources for biobanking. Traditionally, jaguar fibroblast-like cells are frozen in cryovials using slow-freezing protocols. However, this approach necessitates access to ultra-low temperature freezers and requires substantial storage capacity. This study aimed to evaluate the efficacy of a rapid-freezing protocol as a feasible alternative for the freezing of jaguar fibroblasts. Following cultivation to the third passage, cells were frozen using either slow-freezing (SF) or rapid-freezing (RF). For SF, cells were frozen in cryovials (-1 ºC/min), whereas RF involved rapid-freezing (-60 ºC/min) using nitrogen vapor and storage in 0.25 mL straws. Post-thaw, cells were re-cultured to assess recovery. Cell viability was assessed via light microscopy (Trypan blue), fluorescence microscopy (acridine orange + propidium iodide), and flow cytometry (Hoechst 33342 + propidium iodide) immediately post-thaw and upon reaching confluency (> 80%). Fresh culture viability assessed by Trypan blue, acridine orange, and flow cytometry was 98.2 ± 0.3%, 88.7 ± 3.9%, and 94.2 ± 2.2%, respectively. No significant differences in viability were observed between SF and RF immediately post-thaw or after subsequent cultivation. All three assays revealed significant differences (p < 0.05) between cells assessed immediately after thawing and those assessed at confluence. Upon reaching confluence, there was no significant difference (p > 0.05) between fresh and frozen-thawed cells. These findings indicate that RF in straws is a viable alternative to SF for jaguar fibroblast cryopreservation, offering optimized storage density for genetic resource banks.

Keywords

big cats, biobanking, conservation, cryopreservation, felids

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Submitted date:
06/27/2025

Accepted date:
04/13/2026

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