Preserving genetic material in cryogenic conditions presents a viable alternative for the protection of species' gene variability. However, there is an enormous need to establish and test cryopreservation protocols that are suitable for each diverse cell type to guarantee technical success in the long run. Considering this, fibroblasts from jaguar (Panthera onca), oncilla (Leopardus tigrinus), and pampas cat (Leopardus colocolo) were subjected to cell characterization and then cryopreservation in different cryoprotectant solutions (2.5%, 10% dimethyl sulfoxide [DMSO] or CryoSOfree™). Further testing was conducted to determine each solution's performance in preserving cell viability. In culture, a growth curve to assess cellular growth potential showed that exponential proliferation lasts for about the first 50 hours of in vitro culturing, declining in pace afterward. L. colocolo and L. tigrinus presented no difference in cell viability while using 2.5% DMSO protocols. P. onca cells did not present difference on viability for both concentrations of DMSO. Protocols using CryoSOfree resulted in a decreased viability of P. onca fibroblasts. Morphological differences between fibroblasts among the species were noted under bright field microscopy and scanning electron microscopy. L. colocolo and P. onca cells are fusiform, and L. tigrinus are spherical. All cells presented cytoplasmic projections. Transmission electron microscopy revealed vacuoles and secretion granules, indicating intense cell activity after thawing. Differences found in the efficiency of cryopreservation protocols according to the type of cryoprotectant indicate that species react differently to freezing and thawing processes. This research evaluates key aspects of in vitro protocols for cryopreservation of wild animals, which need to be optimized to guarantee successful cell culturing. More suitable protocols lead to increased efficiency in establishing fibroblast cryobanks and also facilitating the use of wild cats' cells in cloning techniques, contributing directly to preserving wild fauna.

中文翻译：

---

细胞特性和冷冻保护剂溶液对三只巴西野猫成纤维细胞活力的影响

在低温条件下保存遗传材料是保护物种基因变异性的可行替代方案。然而，从长远来看，非常需要建立和测试适用于各种不同细胞类型的冷冻保存方案，以保证技术上的成功。考虑到这一点，来自美洲虎 ( Panthera onca )、oncilla ( Leopardus tigrinus ) 和潘帕斯猫 ( Leopardus colocolo ) 的成纤维细胞) 进行细胞表征，然后在不同的冷冻保护剂溶液（2.5%、10% 二甲基亚砜 [DMSO] 或 CryoSOfree™）中冷冻保存。进行了进一步的测试以确定每种溶液在保持细胞活力方面的性能。在培养中，用于评估细胞生长潜力的生长曲线显示，指数增殖持续约前 50 小时的体外培养，之后速度下降。L. colocolo和L. tigrinus在使用 2.5% DMSO 方案时在细胞活力方面没有差异。P. onca细胞在两种 DMSO 浓度下的生存能力没有差异。使用 CryoSOfree 的方案导致P. onca 的生存力降低成纤维细胞。在明视野显微镜和扫描电子显微镜下观察到物种间成纤维细胞之间的形态差异。L. colocolo和P. onca细胞为梭形，L. tigrinus为球形。所有细胞都呈现细胞质突起。透射电子显微镜显示液泡和分泌颗粒，表明解冻后细胞活动强烈。根据冷冻保护剂的类型在冷冻保存协议的效率中发现的差异表明物种对冷冻和解冻过程的反应不同。该研究评估了体外的关键方面野生动物冷冻保存方案，需要对其进行优化以保证细胞培养成功。更合适的方案提高了建立成纤维细胞冷冻库的效率，并促进了在克隆技术中使用野猫的细胞，直接有助于保护野生动物群。