Production of Boer goat (Capra hircus) by nuclear transfer of cultured and cryopreserved fibroblast cells into slaughterhouse-derived oocytes

Abstract

ABstrAct:The present study reports a birth of goat clone produced by nuclear transfer of cultured and cryopreserved fibroblast cells into slaughterhouse-derived oocytes. The donors of somatic cells were fibroblasts derived from the ear skin of a Boer goat while the recipient ooplasm was in vitro matured oocytes of Huanghuai white goat, an Anhui native goat species. The reconstructed embryos were cultured in vitro and then the morphologically normal embryos were transferred to the surrogates. The reconstructed embryos were surgically transferred into 37 recipient surrogates, Huanghuai white goats with natural oestrus. Five of them were treated with hCG after transfer. Among them, one was pregnant and gave birth to a live kid. Due to the improper delivery aid, the cloned kid died accidentally after birth. The cloned kid was then anatomised to observe the viscera development, and the results showed that the organs were normal. Paraffin tissue slices were prepared and stained to ensure the viscera development further, and the results suggested that the organs also developed well in spite of incipient hydropericardium. The microsatellite analysis identified the cloning. It is suggested that the optimised nuclear transfer protocol and proper hCG treatment lead to the successful birth of a goat clone.Keywords: goat; fibroblast; clone; hCG 449Czech J. Anim. Sci., 54, 2009 (10): 448-460 Original Paper are needed to elucidate to what extent the origin of the oocyte affects the development of resultant reconstructed embryos into foetuses and possibly offspring in the goat. The administration of exogenous hormones like human chorionic gonadotropin (hCG) was used in cattle, sheep and goat (Fonseca and Torres, 2005). This hormonal administration can efficiently promote the ovulation of the first-wave dominant follicle forming the accessory corpus luteum.The Boer goat, a famous subspecies originated from South Africa, has advantages primarily for meat production. In addition to its docility and adaptability to various environments and feeds, this breed grows faster and its meat is popular among consumers. Moreover, the Boer goat phenotype including a brown and white head with big ears may be utilised as a marker in genetic studies.The present study was therefore undertaken to initiate Boer goat cloning by SCNT using native goats as oocyte donors and foster mothers to examine whether the hCG injection to recipients after embryo transfer improves the success rate. MAtErIAL AND MEtHODs selection of animalsAn adult Boer goat was used as a donor of somatic cells. Huanghuai white goat, an Anhui native goat species, was used as donors of oocytes as well as recipients of SCNT embryos. More than 70 goats were raised on a farm as recipient candidates, receiving general health care, and their oestrus was also observed. Exclusively healthy individuals were selected for the present experiments. Thirty-seven recipient surrogates were used altogether in the present study.Blood samples were obtained by jugular venipuncture using heparin-vac...