title: Porcine embryo production following in vitro fertilization and intracytoplasmic sperm injection from vitrified immature oocytes matured with a granulosa cell co-culture system.
abstract: This study was designed to evaluate the capacity of vitrified-warmed porcine immature oocytes to mature and to be fertilized using in vitro fertilization or intracytoplasmic sperm injection, and to determine the subsequent embryo development. Immature oocytes were vitrified using ethylene glycol and dimethylsulphoxide as cryoprotectants and the Cryolock method. After warming oocytes were cultured 44 h for maturation. Oocytes were randomly distributed in three treatment groups and subjected to in vitro fertilization (Experiment 1) or intracytoplasmic sperm injection (Experiment 2) procedures. The results indicate that the embryo development was higher in denuded oocytes co-cultured with granulosa cells (NkO-CC group) fertilized by in vitro fertilization or intracytoplasmic sperm injection compared to cumulus-cell oocyte complexes (COCs group), showing no significant differences with control. Vitrified denuded oocytes matured with a co-culture system NkO-CC group, displayed higher cleavage rate and blastocyst production than vitrified COCs group. Blastocysts were successfully obtained after IVF and ICSI procedures; however, the development to the blastocyst stage was better after IVF. These results show that the vitrification-warming media, the employment of a granulosa cell co-culture system and the Cryolock method during vitrification, increased the nuclear and cytoplasmic maturation of vitrified porcine immature oocytes. Further experiments are required to enhance porcine embryo production after vitrification.
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico; Doctorado en Ciencias Biológicas y de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico. 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, 09340 DF, Mexico. 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, 09340 DF, Mexico. Electronic address: anaelenalemus@prodigy.net.mx.
authors.forename: Ana E
authors.initials: AE
authors.lastname: Lemus
authors.affiliation: Departamento de Medicina y Cirugía Animal, Universidad de Murcia, Espinardo 30100, Spain. Electronic address: ccuello@um.es.
authors.forename: Cristina
authors.initials: C
authors.lastname: Cuello
authors.affiliation: Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico. Electronic address: bet@xanum.uam.mx.
authors.forename: Miguel
authors.initials: M
authors.lastname: Betancourt
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002454:Cell Differentiation; D018920:Coculture Techniques; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000494:pharmacology*; D054885:Cumulus Cells / Q000166:cytology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D046149:Embryo Culture Techniques; D047108:Embryonic Development; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000166:cytology*; D006801:Humans; D009865:Oocytes / Q000166:cytology*; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary*; D013552:Swine / Q000502:physiology*; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide
keywords: Cryolock; Embryo development; Granulosa cells; ICSI; IVF; Immature oocytes; Porcine; Vitrification