No synergistic effect of extracellular cryoprotectants with dimethyl sulfoxide in the conservation of northern tiger cat fibroblasts

The success of somatic cell cryobanks is dependent on establishing reproducible cryopreservation methodologies. We supposed that associated extracellular cryoprotectants (sucrose and L-proline) with 2.5 or 10 % dimethyl sulfoxide (Me2SO) could guarantee better northern tiger cat cells quality rates...

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Published in:Cryobiology 2025-03, Vol.118, p.105169, Article 105169
Main Authors: Viana, João Vitor da Silva, Oliveira, Lhara Ricarliany Medeiros de, Rodrigues, Luanna Lorenna Vieira, Moura, Yasmin Beatriz França, Pereira, Antonia Beatriz Mendonça, Alves, Patrícia Vasconcelos, Silva, Herlon Victor Rodrigues, Pereira, Alexsandra Fernandes
Format: Article
Language:English
Subjects:
apoptosis
Biodiversity conservation
cryobiology
Cryopreservation
cryoprotectants
dimethyl sulfoxide
fibroblasts
L-proline
membrane potential
metabolism
mitochondrial membrane
proline
reactive oxygen species
Somatic cell
somatic cells
species
Sucrose
synergism
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Summary:The success of somatic cell cryobanks is dependent on establishing reproducible cryopreservation methodologies. We supposed that associated extracellular cryoprotectants (sucrose and L-proline) with 2.5 or 10 % dimethyl sulfoxide (Me2SO) could guarantee better northern tiger cat cells quality rates after thawing when compared to Me2SO alone. Therefore, we evaluated the effects of sucrose or L-proline with 2.5 or 10 % Me2SO on the cryopreservation of northern tiger cat fibroblasts. Somatic cells were also cryopreserved with 2.5 % or 10 % Me2SO alone. All cells were analyzed for morphology, membrane integrity, proliferative activity, metabolism, apoptosis classification, reactive oxygen species (ROS) levels, and mitochondrial membrane potential (ΔΨm). Regardless of the cryoprotective solution, cryopreservation did not affect morphology, membrane integrity after culture, proliferative activity, and metabolism (P > 0.05). However, immediately after thawing, 2.5 % Me2SO with L-proline and 10 % Me2SO promoted higher rates of membrane integrity when compared to the other cryopreserved groups (P  0.05). However, the percentage of viable cells evaluated by apoptosis classification was reduced when using 10 % Me2SO with L-proline compared to non-cryopreserved groups (P 
ISSN:0011-2240
1090-2392
1090-2392
DOI:10.1016/j.cryobiol.2024.105169