[Invitro Production of Bovine Embryos By Coculture With Oviductal Cells After Invitro Maturation and Fertilization]
(1991) Annales de Medecine Veterinaire (Print Edition) — Vol. 135, n° 5, p. 365-371 (1991)
Files
No attached file found for this publication.
Details
- Authors
- Abstract
- Immature oocytes are harvested from ovaries collected at slaughterhouse. They are matured for 24 hours <<in vitro>> : 82% of them reach the metaphase II stage. They are then fertilized <<in vitro>> using thawed bull semen. The living spermatozoa are selected by centrifugation on a 45 %-90 % Percoll gradient; 10-mu-g/ml heparin are added to the fertilization medium to induce capacitation : 77 % of the oocytes display two pronuclei after this procedure and are considered to be normally fertilized. Zygotes are then cultured <<in vitro>> . In a first trial we compared the results of a seven days culture either (l) in droplets of 199 medium, or (2) in the same medium conditioned (MC) by means of bovine oviduct epithelial cells (BOEC), or (3) by the combination of 4 days in MC followed by 3 days on a monolayer of BOEC. For these three culture conditions we obtained respectively 4%, 8% and 18% of expanded blastocysts as compared to the number of immature oocytes collected. The second trial did use the protocol (3) in view to compare bovine (BOEC) with rabbit (ROEC) oviduct epithelial cells. The global results were poorer than above, however we observed a higher number of expanded blastocysts using BOEC (8,6%) than with ROEC (2,5%). In some more recent trials using 4 days culture in MC followed by 3 days on the BOEC monolayer we did obtain 101 expanded blastocysts from 468 immature oocytes (22%). Six cows were used as foster mothers receiving from 1 to 4 embryos. Four of them became pregnant and gave birth to 4 normal calves. This result allows us to conclude that <<in vitro>> produced embryos, at least some of them, are able to produce a normal pregnancy in cattle.
- Affiliations
Citations
Boccart, C., Mermillod, P., Massip, A., Delecoeuillerie, C., & Dessy, F. (1991). [Invitro Production of Bovine Embryos By Coculture With Oviductal Cells After Invitro Maturation and Fertilization]. Annales de Medecine Veterinaire (Print Edition), 135(5), 365-371. https://hdl.handle.net/2078.5/137498 (Original work published 1991)