In vitro canine oocyte nuclear maturation in homologous oviductal cell co-culture with hormone-supplemented media.
The aim of the present research was to verify the influence of oviductal cell co-culture previously supplemented with steroids (estrogen, progesterone, or both) on IVM rates for oocytes from anestrous bitches that were cultured in vitro for 48, 72 and 96 h. Oocytes harvested from anestrous bitches were selected and allocated into four groups: Group 1 (co-culture in oviductal epithelial cells without hormonal supplementation-control); Group 2 (estrogen supplementation); Group 3 (progesterone supplementation); Group 4 (estrogen+progesterone supplementation). The oviductal epithelial cell culture was established 72 h prior to oocyte co-culture. After periods of 48, 72 and 96 h, the degree of oocyte nuclear maturation was assessed. Co-culture in oviductal epithelial cells with estrogen was not as beneficial for canine IVM as supplementation with progesterone and estrogen, or progesterone supplementation alone. Therefore, it was feasible to use co-culture with oviductal epithelial cells obtained from anestrous bitches for IVM (monolayer culture with oviduct cells previously supplemented with progesterone). Final stages of oocyte maturation were achieved at 72 and 96 h of culture; therefore, the duration of maturation for oocytes obtained from bitches in different stages of the estrous cycle should be taken into account.
Department of Animal Reproduction, Faculty of Veterinary Medicine, University of São Paulo, Av. Prof. Orlando Marques de Paiva, 87, Cidade Universitária, 05508-000 São Paulo, SP, Brazil. cacavann@usp.br
Camila Infantosi
CI
Vannucchi
Clair Motos
CM
de Oliveira
Mariana Groke
MG
Marques
Mayra Elena Ortiz Avila
ME
Assumpção
José Antonio
JA
Visintin
D000818:Animals; D001562:Benzimidazoles / Q000737:chemistry; D002467:Cell Nucleus / Q000502:physiology; D018920:Coculture Techniques / Q000662:veterinary; D003470:Culture Media; D004285:Dogs / Q000502:physiology*; D004847:Epithelial Cells / Q000166:cytology; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000662:veterinary; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D011374:Progesterone / Q000494:pharmacology*
D001562:Benzimidazoles; D003470:Culture Media; D005456:Fluorescent Dyes; D011374:Progesterone; D004958:Estradiol; C017807:bisbenzimide ethoxide trihydrochloride