Clone of Chinese Jinan red-cross yellow cattle and evaluation of reproductive characteristics of cloned calf

Somatic cell clone technology is a viable approach to preserving endangered livestock and wildlife genetic resources. In the present research, somatic cell nuclear transfer （SCNT） was performed using granulose cells from the critical endangered Chinese red-cross yellow cattle as donor cells. A total of 211 oocytes were manipulated and 166 （79%） of them were successfully enucleated. 112 （67.4%） SCNT embryos were reconstructed, 94 （83%） of them cleaved, and 48 （43 %） of them developed to blastocyst stage. SCNT blastocysts were transferred to 6 Holstein recipients, and 2 （33%） of them were found to be pregnant. One of them maintained to term and delivered a calf, whereas another aborted. Effect of different fusion buffer （mannitol vs. Zimmerman fusion buffer） and different activation methods （calcium ionophore＋6-DMAP vs. cycloheximide＋CB） on fusion rate and development of SCNT embryos were investigated. The results indicated that： （i） on condition of two DC pulses of 2.5 kV/cm for 10 μs each, fusion rates were higher in mannitol solution than in Zimmerman fusion buffer （71% vs. 61%, respectively, p 〈 0.05）, but the blastocysts rates did not differ between two treatments （36 % vs. 39 %, p〉0.05 ）; （ii） There was no significant difference in development rates to the blastocyst stage for SCNT embryos activated by calcium ionophore＋6-DMAP or by cycloheximide＋CB （42% vs. 46%, respectively, p〉0.05）. Microsatellite DNA analysis examining 28 loci confirmed that the cloned calf was genetically identical to the donor Jinan red-cross yellow cattle and different from the recipient females. Growth and reproductive performance of cloned cow were evaluated, and there were no difference i cross-red n it between cloned and normal control Jinan yellow cattle. Furthermore, the cloned yellow cow has delivered a healthy yellow calf.

Clone of Chinese Jinan redcross yellow cattle and evaluation of reproductive characteristics of cloned calf

Somatic cell clone technology is a viable approach to preserving endangered livestock and wildlife genetic resources. In the present research, somatic cell nuclear transfer (SCNT) was performed using granulose cells from the critical endangered Chinese red-cross yellow cattle as donor cells. A total of 211 oocytes were manipulated and 166 (79%) of them were successfully enucleated. 112 (67.4%) SCNT embryos were reconstructed, 94 (83%) of them cleaved, and 48 (43%) of them developed to blastocyst stage. SCNT blastocysts were transferred to 6 Holstein recipients, and 2 (33%) of them were found to be pregnant. One of them maintained to term and delivered a calf, whereas another aborted. Effect of different fusion buffer (mannitol vs. Zimmerman fusion buffer) and different activation methods (calcium ionophore+6-DMAP vs. cycloheximide+CB) on fusion rate and development of SCNT embryos were investigated. The results indicated that: (i) on condition of two DC pulses of 2.5 kV/cm for 10 μs each, fusion rates were higher in mannitol solution than in Zimmerman fusion buffer (71% vs. 61%, respectively, p<0.05=, but the blastocysts rates did not differ between two treatments (36% vs. 39%, p>0.05); (ii) There was no significant difference in development rates to the blastocyst stage for SCNT embryos activated by calcium ionophore+6-DMAP or by cycloheximide+CB (42% vs. 46%, respectively, p>0.05). Microsatellite DNA analysis examining 28 loci confirmed that the cloned calf was genetically identical to the donor Jinan red-cross yellow cattle and different from the recipient females. Growth and reproductive performance of cloned cow were evaluated, and there were no differencei cross-red n it between cloned and normal control Jinan yellow cattle. Furthermore, the cloned yellow cow has delivered a healthy yellow calf.