Co-culture with granulosa cells improve the in vitro maturation ability of porcine immature oocytes vitrified with cryolock
Autor: | Eduardo Casas, Ana E. Lemus, Miguel Betancourt, Mario Teteltitla-Silvestre, Fahiel Casillas, Zayil Salazar, Yvonne Ducolomb |
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Rok vydání: | 2014 |
Předmět: |
Ethylene Glycol
Cryoprotectant Cell Survival Swine In Vitro Oocyte Maturation Techniques Biology General Biochemistry Genetics and Molecular Biology Cryopreservation Andrology chemistry.chemical_compound Cryoprotective Agents medicine Animals Vitrification Dimethyl Sulfoxide Cells Cultured Granulosa Cells Dimethyl sulfoxide Trehalose General Medicine Oocyte Coculture Techniques In vitro maturation medicine.anatomical_structure chemistry Immunology Oocytes Female General Agricultural and Biological Sciences |
Zdroj: | Cryobiology. 69(2) |
ISSN: | 1090-2392 |
Popis: | This study was designed to evaluate the efficiency of two oocyte vitrification-warming procedures using two different devices: Superfine Open Pulled Straws (SOPS) and Cryolock, as well as the effect of the co-culture of vitrified immature oocytes with fresh granulosa cells to improve in vitro maturation (IVM). Immature oocytes were vitrified with two procedures: A) Oocytes were exposed to an increasing concentration of ethylene glycol (EG) from 4% to 35% with 0.5 M trehalose. They then, were loaded in SOPS or Cryolock. For warming, oocytes were exposed to decreasing concentrations of trehalose 0.3, 0.2 and 0.1 M for IVM. B) Oocytes were exposed to two mixtures of EG and dimethylsulfoxide (Me2SO), at 7.5% and 16%, both with 0.4 M of sucrose and then loaded in SOPS or Cryolock and stored in liquid nitrogen. For warming, oocytes were exposed to a single concentration of sucrose 0.5M. After warming, viability was determined; and after 44 h of IVM both viability and meiotic stages were evaluated. The results indicate no significant differences between procedures A and B with SOPS in all maturation stages, reaching a maximum maturation rate of 21%. As to Cryolock, significant differences were observed between both procedures, being procedure B, more efficient with a yield of 38% in MII stage and increased to 49% due to the co-culture with fresh granulosa cells. In conclusion, viability and maturation rates were improved with Cryolock and procedure B with the co-culture system in vitrified immature oocytes. |
Databáze: | OpenAIRE |
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