Cholesterol added prior to vitrification on the cryotolerance of immature and in vitro matured bovine oocytes
Arcarons Deseures, Núria (Universitat Autònoma de Barcelona. Departament de Medicina i Cirurgia Animals)
Morató, Roser (Universitat Autònoma de Barcelona. Departament de Medicina i Cirurgia Animals)
Vendrell-Flotats, Meritxell (Universitat Autònoma de Barcelona. Departament de Medicina i Cirurgia Animals)
Yeste Oliveras, Marc (Universitat de Girona. Departament de Biologia)
López Béjar, Manel (Universitat Autònoma de Barcelona. Departament de Medicina i Cirurgia Animals)
Rajapaksha, Kosala (University of Saskatchewan. Development Centre and Western College of Veterinary Medicine)
Anzar, Muhammad (University of Saskatchewan. Development Centre and Western College of Veterinary Medicine)
Mogas Amorós, Teresa, dir. (Universitat Autònoma de Barcelona. Departament de Medicina i Cirurgia Animals)

Fecha:	2017
Resumen:	This study examines whether incorporating cholesterol-loaded methyl-β-cyclodextrin (CLC) in the bovine oocyte plasma membrane improves oocyte tolerance to vitrification. In vitro matured oocytes were incubated with 2 mg/ml BODIPY-labeled CLC for different time intervals in FCS or PVA supplemented medium or exposed to different CLC concentrations to examine the subcellular localization of cholesterol by confocal microscopy live-cell imaging. Subsequently, the effects of optimized CLC concentrations and incubation times prior to vitrification on early embryo development were assessed. Then, we evaluated the effects of pretreatment with 2 mg/ml CLC for 30 min before the vitrification of immature (GV) and in vitro matured (MII) oocytes on developmental competence and gene expression. Our results indicate a high plasma membrane labeling intensity after 30 min of incubation with 2 mg/ml CLC for 30 min, regardless of the holding medium used. When oocytes were incubated with 1 mg/ml, 2 mg/ml and 3 mg/ml of CLC, intense labeling was observed at the plasma membrane after 40, 30 and 20 min, respectively. CLC pre-treatment before the vitrification of bovine oocytes did not affect subsequent cleavage and embryo development rates irrespective of CLC concentrations, incubation times or meiotic stage. However, pretreatment seems to improve the quality of embryos derived from vitrified oocytes, mainly when oocytes were vitrified at the GV stage.
Ayudas:	Ministerio de Economía y Competitividad AGL2013-46769-P
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Colesterol ; Fecundació in vitro
Publicado en:	PloS one, september 2017, ISSN 1932-6203

DOI: 10.1371/journal.pone.0184714
PMID: 28910399

21 p, 8.5 MB

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