In vitro development of DNA-injected embryos co-cultured with goat oviduct epithelial cells in Korean native goats (Capra hircus aegagrus)

Abstract

In vitro development of Korean native goat embryos was investigated in 2 different culture systems with and without goat oviduct epithelial cells (GOEC), Estrus was synchronized by inserting intravaginal progestagen- impregnated sponge (Veramix) containing 60 mg medroxyprogesterone acetate (MAP) for 14 d. Superovulation was induced with follicle stimulating hormone (FSH). Goat ova were surgically obtained by retrograde flushing the oviducts of does at 66 to 68 h after MAP removal. Mean number of recovered ova per doe was 7.28 ± 3.91, and the proportion of fertilized embryos in recovered ova was 66.5% (121/182). Fertilized embryos were cultured for 9 d in CR1aa medium supplemented with 10% estrous goat nun (EGS) at 38.5°C, 5% CO2 in air. There was no difference in development of the embryos to the morula stage between the 2 culture systems (84.4 and 84.0%, respectively). However, developmental rate to blastocysts (65.6%) of the embryos co-cultured with GOEC was significantly higher than of those (12.0%) cultured without GOEC (P<0.001). Goat zygotes were injected with bovine β-casein/human lactoferrin cDNA fusion gene (pBL1). When the DNA-injected embryos were co-cultured with GOEC, developmental rates of the embryos to the morula and blastocyst stages were 82.9 and 36.6%, respectively. The results obtained in this study indicate that 'blocking' of in vitro development of Korean native goat embryos appears to occur at the morula stage, but can be overcome to some extent by co-culture with GOEC. In the co-culture system, DNA-injected goat embryos could successfully develop to normal hatching blastocysts.