Ultrarapid Vitrification of Snow Leopard (Panthera uncia) Fibroblasts

Dinmukhamed Toishybek, Grachev Alexey, Maxim Bespalov, M. Salmenova, Bibigul Katubayeva, Indira Kumayeva, N. Bizhanova, Yelena Assanova, Yerzhan Toishibekov, M.Y. Salmenova

2026

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DOI 10.30919/ee2558
Data confidence 85 high
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Abstract

Somatic cell biobanking supports conservation of endangered felids, yet cryopreservation protocols for the snow leopard (Panthera uncia) remain insufficiently standardised.We evaluated the effects of vitrification-solution composition and cooling procedure on post-thaw membrane integrity of dermal fibroblasts from 16 wild individuals.Cells were assigned to six treatments comprising 20% dimethyl sulphoxide (DMSO), 20% ethylene glycol (EG), or 20% DMSO + 20% EG, each with 0.5 M sucrose, combined with conventional cooling in liquid nitrogen (-196 °C) or VitMaster-assisted supercooled vitrification (-205 °C).Cryoprotectant composition strongly affected post-thaw membrane integrity (F2, 90 = 419.70,p < 0.0001, partial η 2 = 0.903).DMSO-EG yielded higher integrity (64.5 ± 0.6%) than DMSO (42.8 ± 0.6%) or EG (43.9 ± 0.6%; both p < 0.0001), whereas DMSO and EG did not differ (p = 0.397).Cooling procedure had no significant effect (F1, 90 = 2.87, p = 0.094, partial η 2 = 0.031), nor did its interaction with cryoprotectant composition (F2, 90 = 0.18, p = 0.837).All treatments reduced membrane integrity.DMSO-EG with conventional cooling yielded the highest integrity, whereas supercooling conferred no demonstrated benefit.Concentration-matched controls are needed to distinguish formulation from total cryoprotectant concentration effects.