Cryotops versus open-pulled straws (OPS) as carriers for the cryopreservation of bovine oocytes: effects on spindle and chromosome configuration and embryo development.
Two experiments were designed to assess the effectiveness of cryopreserving bovine MII oocytes using cryotops as the carrier system for vitrification. In the first experiment, we examined the developmental competence of oocytes after: (i) vitrification in open-pulled straws (OPS method); or (ii) vitrification in <0.1mul medium droplet on the surface of a specially constructed fine polypropylene strip attached to a plastic handle (Cryotop method). In the second experiment, warmed oocytes that had been vitrified in OPS or cryotops were fixed to analyze spindle and chromosome configuration. In all experiments both cow and calf oocytes were used. Significantly different fertilization rates were observed between the vitrification groups: 31.5% and 20.2% for the cow and calf oocytes vitrified in OPS, respectively, versus 46.1% and 46.4% for the oocytes vitrified using cryotops. After in vitro fertilization, 3.8% of the calf oocytes and 5.3% of the cow oocytes developed to the blastocyst stage. All blastocysts from vitrified oocytes resulted from the Cryotop method. A significantly lower percentage of the OPS-vitrified calf oocytes showed a normal spindle configuration (37.8%) compared to control fresh oocytes (69.9%), while normal spindle and chromosome configurations were observed in a significantly higher proportion of the cryotop-vitrified calf oocytes (60.2%). For the cow oocytes, 60.6% in the OPS group and 60.3% in the Cryotop group exhibited a normal morphology after warming. These findings suggest the cryotop system is a more efficient carrier for vitrification than OPS for the cryopreservation of bovine oocytes.
Morató
Roser
R
Departament de Medicina i Cirurgia Animals, Universitat Autònoma de Barcelona, Facultat de Veterinaria, Edifici V, E-08193 Bellaterra, Barcelona, Spain.
Izquierdo
Dolors
D
Paramio
Maria Teresa
MT
Mogas
Teresa
T
D000818:Animals; D001755:Blastocyst / Q000187:drug effects; D002417:Cattle; D018929:Cell Culture Techniques; D002470:Cell Survival; D033481:Chromosomes, Mammalian / Q000187:drug effects / Q000648:ultrastructure; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro; D047569:Laser Scanning Cytometry; D008297:Male; D008968:Molecular Conformation; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development / Q000502:physiology*; D017239:Paclitaxel / Q000494:pharmacology
D003451:Cryoprotective Agents; D017239:Paclitaxel