Naslov
The current status of in vitro bovine embryo production in the republic of Croatia

Autori
Getz, Iva ; Matković, Mario ; Makek, Zdenko ; Tomašković Antun ; Cergolj, Marijan ; Dobranić, Tomislav ; Samardžija, Marko

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, cjeloviti rad (in extenso), pregledni

Izvornik
Proceedings of the II. Middle-European Congress for Buiatrics "Effect of Cattle-Herd Health on Production Efficiency" / Kovač, Gabriel ; Maraček, Imrich - Košice : Dunadan, 2000, 92-95

Skup
II. Middle-European Congress for Buiatrics "Effect of Cattle-Herd Health on Production Efficiency"

Mjesto i datum
Vysoké Tatry, Slovačka, 11.05.2000. - 13.05.2000

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
bovine; oocyte maturation; in vitro fertilization; cooculture; granulosa cells; embryo
(in vitro fertilization; cooculture; granulosa cells; embryo)

Sažetak
Since the first calf as a result of in vitro fertilization was born in 1982, the in vitro technology has advanced enormously. In vitro production (IVP) of bovine embryos has become a routine procedure in many laboratories. It is now in the process of transition from strictly scientific studies to a technology which permits breeding plans and implementation of other embryotechnologies such as freezing, twining, cloning, transgenesis, sexing of embryos and others. Therefore, three years ago we started a project "In vitro fertilization in cows", with the aim to establish the basic protocol for in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC) of bovine oocytes and to introduce this new biotechnology in our veterinary medicine and breeding programs. Cumulus oocyte complexes (COCs) were collected from ovaries of slaughtered cows by aspiration of antral follicles with diameter less than 8 mm. A total of 426 oocytes were selected for IVM from 158 ovaries (2.7 oocytes per ovary, i.e. 5.4 per pair). The oocytes were matured in drops of maturation medium (TCM 199 medium supplemented with fetal calf serum and hormones). After 24 hours of incubation at 39oC and with 5% CO2, matured oocytes were inseminated in vitro with frozen/thawed bull semen, prepared by gradient separation. Forty-eight hours after insemination 4- and 8-cell embryos were selected and cultured in vitro to the blastocyst stage in supplemented TCM 199 medium in co-culture with granulosa cells. Maturation rate was determined based on expansion of cumulus and corona cells and it was 87.6%. Fertilization i.e. cleavage rate determined 48 hours after insemination was 56.3% and blastocysts production rate observed on day 7 was 14.7%. Bovine blastocysts produced in vitro were of good quality, according to IETS standards and suitable for embryo transfer or cryopreservation. The next programmed step is the ultrasound-guided transvaginal ovum pick-up (OPU) from the ovaries of living cows for embryo transfer purposes.

Izvorni jezik
Engleski

POVEZANOST RADA