Fish (Oncorhynchus mykiss) spermatozoa cryoprotectant-free vitrification: Stability of mitochondrion as criterion of effectiveness

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datacite.alternateIdentifier.citationAnimal Reproduction Science, Vol. 124, 1-2, 125-131, 2011es
datacite.alternateIdentifier.doi10.1016/j.anireprosci.2011.02.023es
datacite.creatorMerino, O.
datacite.creatorRisopatrón, J.
datacite.creatorSánchez, R.
datacite.creatorIsachenko, E.
datacite.creatorFigueroa Villalobos, Elías
datacite.creatorValdebenito Isler, Iván
datacite.creatorIsachenko, V.
datacite.date2011
datacite.date.issued2012-02-08
datacite.subjectCriopreservaciónes
datacite.subjectVitrificaciónes
datacite.subjectEspermatozoideses
datacite.subjectTrucha arcoírises
datacite.titleFish (Oncorhynchus mykiss) spermatozoa cryoprotectant-free vitrification: Stability of mitochondrion as criterion of effectivenesses
dc.date.accessioned2012-02-08T16:50:53Z
dc.date.available2012-02-08T16:50:53Z
dc.description.abstractThe aim of the present investigations was to test a novel technology comprising cryoprotectant-free vitrification of the spermatozoa of rainbow trout and to study the ability of sucrose and components of seminal plasma to protect these cells from cryoinjuries. Spermatozoa were isolated and vitrified using five different mediums: Group 1: standard buffer for fish spermatozoa, Cortland®-medium (CM, control); Group 2: CM+1% bovine serum albumin (BSA); Group 3: CM+1% BSA+0.125M sucrose; Group 4: CM+1% BSA+40% seminal plasma; and Group 5: CM+1% BSA+40% seminal plasma+0.125M sucrose. For cooling, 20μL suspensions of cells from each group were dropped directly into liquid nitrogen. For warming, the spheres containing the cells were quickly submerged in CM+1% BSA at 37°C with gentle agitation. The quality of spermatozoa before and after vitrification was analysed by the evaluation of motility, cytoplasmic membrane integrity (SYBR-14/propidium iodide staining technique), and mitochondrial membrane integrity (JC-1 staining). Motility (86%, 71%, 80%, 81%, and 82%, for Groups 1, 2, 3, 4, and 5, respectively) and cytoplasmic membrane integrity (90%, 82%, 83%, 84%, and 87%, respectively) of spermatozoa in all the 5 groups were not decreased significantly. All tested solutions can be used for vitrification of fish spermatozoa with good post-warming motility and cytoplasmic membrane integrity. However, mitochondrial membrane potentials of the spermatozoa in Groups 1, 2, 3, 4, and 5 were changed significantly (6%, 50%, 37%, 55%, and 34%, respectively) (P1,2,3,4,5<0.001; P2,3,4,5 <0.01)(P3-5>0.1). This rate was maximal in Group 4 (CM+1% BSA+40% seminal plasma). In conclusion, this is the first report about successful cryoprotectant-free cryopreservation of fish spermatozoa by direct plunging into liquid nitrogen (vitrification). Vitrification of fish spermatozoa without permeable cryoprotectants is a prospective direction for investigations: these cells can be successfully vitrified with 1% BSA+40% seminal plasma without significant loss of important physiological parameters. © 2011 Elsevier B.V.es
dc.formatPDFes
dc.identifier.urihttps://repositoriodigital.uct.cl/handle/10925/595
dc.language.isoenes
dc.sourceAnimal Reproduction Sciencees
oaire.resourceTypeArtículo de Revistaes
uct.carreraIngeniería en Acuiculturaes
uct.catalogadorpopes
uct.comunidadRecursos Naturaleses
uct.facultadFacultad de Recursos Naturaleses
uct.indizacionISIes
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