title: Co-culture with granulosa cells improve the in vitro maturation ability of porcine immature oocytes vitrified with cryolock.
abstract: 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.
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México; Maestría en Biología de la Reproducción Animal, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa Av. San Rafael Atlixco 186, 09340 DF, México. Electronic address: fahiel.ca@gmail.com.
authors.forename: Fahiel
authors.initials: F
authors.lastname: Casillas
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México; Maestría en Biología de la Reproducción Animal, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa Av. San Rafael Atlixco 186, 09340 DF, México. Electronic address: teteltitla7@gmail.com.
authors.forename: Mario
authors.initials: M
authors.lastname: Teteltitla-Silvestre
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México. Electronic address: duco@xanum.uam.mx.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Ducolomb
authors.affiliation: Departamento de Biología de la Reproducción, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México. Electronic address: anaelenalemus@prodigy.net.mx.
authors.forename: Ana E
authors.initials: AE
authors.lastname: Lemus
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México. Electronic address: zayil.salazar@gmail.com.
authors.forename: Zayil
authors.initials: Z
authors.lastname: Salazar
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México. Electronic address: dino@xanum.uam.mx.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Casas
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México. Electronic address: bet@xanum.uam.mx.
authors.forename: Miguel
authors.initials: M
authors.lastname: Betancourt
mesh_terms: D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods / Q000662:veterinary*; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000378:metabolism; D019855:Ethylene Glycol / Q000378:metabolism; D005260:Female; D006107:Granulosa Cells / Q000166:cytology*; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology*; D013552:Swine; D014199:Trehalose / Q000378:metabolism; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D014199:Trehalose; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide
keywords: Cryolock; Cryoprotectants; Granulosa cells co-culture; IVM; Immature oocytes; Porcine; SOPS; Vitrification