RESUMEN
BACKGROUND: The synergistic action among the different extracellular cryoprotectants could improve somatic cell quality after thawing and provide bases for the formation of biobanks for red-rumped agoutis. OBJECTIVE: This study evaluated the interactions among sucrose (SUC) and concentrations of serum fetal bovine (FBS) on the cryopreservation of somatic cells derived from red-rumped agoutis. MATERIALS AND METHODS: Cells were cryopreserved with 10% dimethyl sulfoxide and different concentrations of FBS (10%, 40%, and 90%) with or without 0.2 M SUC, totaling six comparison groups. Non-cryopreserved cells were used as a control. Cells were evaluated for viability, metabolic activity, proliferative activity, reactive oxygen species (ROS), mitochondrial membrane potential and apoptosis levels. RESULTS: No difference was observed among cryopreserved with DMSO containing (10FBS, 10FBS-SUC, 40FBS, 40FBS-SUC, 90FBS, 90FBS-SUC) and non-cryopreserved groups for viability, metabolic activity, proliferative activity, and ROS levels. Interestingly, only cells cryopreserved with 90% FBS and SUC maintained the mitochondrial membrane potential like the control. This indicates that at high concentrations of FBS, SUC contributes to the maintenance of this parameter in cryopreserved cells. Moreover, at concentrations of 10% and 40% of FBS, SUC contributed to the maintenance of viability evaluated by the levels of apoptosis evaluated after thawing. In summary, we verified that 90% FBS and 0.2 M SUC promote greater ability of cells after thawing. Additionally, SUC positively acts in cryopreservation solutions containing 10% and 40% FBS. CONCLUSION: This information is essential to an understanding of the mechanisms involved in the interactions of extracellular cryoprotectants in somatic cell cryopreservation solutions of red-rumped agoutis. DOI: 10.54680/fr23210110212.
Asunto(s)
Criopreservación , Dasyproctidae , Animales , Bovinos , Criopreservación/veterinaria , Sacarosa/farmacología , Especies Reactivas de Oxígeno , Crioprotectores/farmacología , Dimetilsulfóxido/farmacología , Supervivencia CelularRESUMEN
BACKGROUND: Skin cryobanks represent important tools for the conservation of the maximum genetic representation of a population, especially those with a certain degree of threat to extinction, such as the ocelot. A relevant step towards the proper establishment of these banks is the definition of adequate cryopreservation techniques for the conservation of the skin. OBJECTIVE: We evaluated the effects of two different techniques [direct vitrification in cryovials (DVC) and solid-surface vitrification (SSV)] for the preservation of ear skin derived from ocelot. MATERIALS & METHODS: For both techniques, we vitrified the ear skin using Dulbeccos modified Eagles medium with 3.0 M dimethyl sulfoxide, 0.25 M sucrose, and 10% fetal bovine serum. Non-cryopreserved tissues were used as control (control group). All tissues were analyzed for their morphometric characteristics by conventional histology and morphological / functional analysis by cell ability during the culture. RESULTS: While tissues cryopreserved by DVC showed similar values for dermis thickness and number of perinuclear halos to the control, tissues cryopreserved by SSV showed similarities to the control regarding the number of melanocytes, percentage of collagen fibers, and numbers of viable cells by apoptosis analysis. Additionally, none of the vitrification techniques affected stratum corneum thickness, number of keratinocytes, tissue proliferative activity, cell viability, or metabolism. CONCLUSION: Both vitrification techniques (DVC and SSV) can be used for the conservation of ocelot skin; however, SSV guarantees a higher cellular quality after in vitro tissue culture in most of the parameters evaluated, such as viability, metabolism, and apoptosis analysis. doi.org/10.54680/fr23110110412.