Desarrollo embrionario de bovino in vitro cocultivado con células oviductales y del cumulus oophorus.

In vitro-fertilized bovine embryos co-cultured with oviductal and cumulus oophorus cells.

BLASTOCYST; SOMATIC cells

La presente investigación tuvo como objetivo evaluar el efecto del cocultivo de los embriones con células oviductales y del cúmulo oóforo (cumulus oophorus) sobre el desarrollo de embriones in vitro en estadio de blastocisto. Los oviductos y ovarios se colectaron de hembras bovinas sacrificadas en un rastro. Las células oviductales se obtuvieron tras una incisión longitudinal del oviducto; las células del cumulo oophorus se colectaron del líquido folicular. Ambos tipos celulares se colocaron para su cultivo en TCM-199 con 20% de Suero Fetal Bovino (SFB). Los complejos cúmulo-ovocitos (COC's) fueron colectados por aspiración de folículos de 2 a 7 mm de diámetro y colocados para su maduración en TCM-199 con 10% de SFB; posteriormente, se fertilizaron utilizando una concentración de 1x106 espermatozoides/mL. Después de la fertilización se dividieron en tres tratamientos: 1) Control, 2) Cocultivo con células de cúmulo, 3) Cocultivo con células del oviducto. El desarrollo se evaluó a 192 horas de cultivo in vitro, determinando la cantidad de embriones divididos, mórulas y blastocistos. Se cultivaron en medio SOF secuencial, por tratamiento: 1) 538 ovocitos, 2) 627 ovocitos y 3) 478 ovocitos. Los resultados fueron los siguientes: 14% de blastocistos en el cocultivo de células cúmulo, 7% en el cocultivo de oviducto y 4% en el control, con una diferencia estadística significativa (p<0,05) entre los tres tratamientos. Se concluye que el uso de medios condicionados con células somáticas puede estimular el desarrollo in vitro hasta estadio de blastocisto de los embriones bovinos. [ABSTRACT FROM AUTHOR]

The research was aimed at evaluating the effect of embryos co-cultured with oviductal and cumulus oophorus cells on the development of blastocyst in vitro. Oviducts and ovaries were collected from slaughtered bovine females. Oviductal cells were obtained after a longitudinal incision of the oviduct and cumulus cells were collected from follicular fluid; both cell types were placed for culture in TCM-199 with 20% Fetal Bovine Serum (FBS). The cumulus-oocyte complexes (COC's) were collected by the aspiration of follicles from 2 to 7 mm diameter and they were placed for maturation in TCM-199 with 10% FBS. Subsequently, they were fertilized using a concentration of 1x106 spermatozoa/mL. After fertilization, they were divided into three treatments: 1) Control, 2) Coculture with cumulus cells and 3) Co-culture with oviduct cells. The development was evaluated at 192 hours of in vitro culture, determining the amount of divided embryos, morulas and blastocysts. They were cultured in sequential SOF medium by the treatment: 1) 538 oocytes, 2) 627 oocytes and 3) 478 oocytes. The results were the following: 14% of blastocysts in the co-culture of cumulus cells, 7% in the co-culture of oviduct and 4% in the control, with a significant statistical difference (p<0.05) among the three treatments. It is concluded that the use of somatic cells in conditioned medium can stimulate the blastocyst stage development in vitro to the bovine embryos. [ABSTRACT FROM AUTHOR]

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