Program optimization before enucleation on ovine somatic cell nuclear transfer.

Yanhua Guo; Yiyuan Zhang; Limin Wang; Hong Tang; Yingli LI; Ping Zhou

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Program optimization before enucleation on ovine somatic cell nuclear transfer.

Author: Yanhua GUO ¹ ; Yiyuan ZHANG ¹ ; Limin WANG ¹ ; Hong TANG ¹ ; Yingli LI ² ; Ping ZHOU ¹
Author Information

1. Institute of Animal Husbandry and Veterinary, Xinjiang Academy of Agricultural and Reclamation Sciences, State Key Laboratory of Sheep Genetic Improvement and Healthy Production, Shihezi 832000, Xinjiang, China.
2. Animal and Medical Quarantine Station of Shihezi, Shihezi 832000, Xinjiang, China.
Publication Type:Journal Article
Keywords: adherent rate; clone sheep offspring; enuclation time; exposure duration of ovaries; nuclear transfer
From: Chinese Journal of Biotechnology 2017;33(5):730-742
CountryChina
Language:Chinese
Abstract: Ovine somatic cell nuclear transfer (SCNT) efficiency remains lower. Therefore, we optimized the program before oocyte enucleation on ovine SCNT. Four experiments were done including exposure duration of ovaries (3 h or 3 to 5 h), duration of oocytes maturation (18 h and 24 h), rate of donor adherent and enucleation time of maturate oocyte. The maturation rates of oocyte, fusion rates and cleavation rates of cloned embryos were used to assess the efficiency of different procedures. The maturation rates of ovaries with 3 h exposure was higher than that of 3 to 5 h (60.18% vs 52.50%) (P<0.05). Embryonic development competence had no significant difference (P>0.05). The maturation rates were significantly different between group18 h and 24 h (53.81% vs 89.06%, P<0.01). Embryonic development competence had no significant difference (P>0.05); fusion rates of donor adherent 30% group was higher than that of 10% group. Embryonic development competence had no significant difference (P>0.05). Different adherent donor characterizes the difference in plateau phase. The cleavation rates of 18 hpm group was higher than that of 16 hpm group. Embryonic development competence had no significant difference (P>0.05), the enucleation of 16 hpm group obtained one clone fetus, we got four clone fetus to repeat the 16 hpm group. Five microsatellite was analyzed by PAGE, the bands indicated that fingerprint of cloned fetus were completely the same as those of donor cells. Our data therefore suggests program optimization before enucleation assurance quality of material which be able to improve the quantity and quality of clone embryos, and optimized scheme can obtain clone sheep offspring.