Cloning of Asian yellow goat (C. hircus) by somatic cell nuclear transfer: Telophase enucleation combined with whole cell intracytoplasmic injection

• Published in: Molecular reproduction and development Vol. 74; no. 1; pp. 28 - 34
• Main Authors: Chen, Da-Yuan, Jiang, Man-Xi, Zhao, Zhen-Jun, Wang, Hai-Long, Sun, Qing-Yuan, Zhang, Li-Sheng, Li, Rui-Chang, Cao, Heng-Hua, Zhang, Quan-Jun, Ma, Dong-Lian
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.01.2007; Wiley-Liss
• Subjects: Animals; Asian yellow goat (C. hircus); Biological and medical sciences; Cell Nucleus - metabolism; Cloning, Organism - methods; Cytoplasm - metabolism; Early stages. Segmentation. Gastrulation. Neurulation; Embryology: invertebrates and vertebrates. Teratology; Fundamental and applied biological sciences. Psychology; Goats - genetics; Microinjections; Nuclear Transfer Techniques; Oocytes - metabolism; somatic cell nuclear transfer; subzonal injection; Telophase; telophase ooplasm; whole cell intracytoplasmic injection; Animal cloning; telophase ooplasm; Cell nucleus; Transgenic animal; Somatic cell; somatic cell nuclear transfer; Vertebrata; Mammalia; Telophase; subzonal injection; Transfer; Goat; Artiodactyla; Ungulata; Asian yellow goat (C. hircus); whole cell intracytoplasmic injection
• ISSN: 1040-452X; 1098-2795
• DOI: 10.1002/mrd.20600

Our and other previous studies have shown that telophase enucleation is an efficient method for preparing recipient cytoplasts in nuclear transfer. Conventional methods of somatic cell nuclear transfer either by electro‐fusion or direct nucleus injection have very low efficiency in animal somatic cell cloning. To simplify the manipulation procedure and increase the efficiency of somatic cell nuclear transfer, this study was designed to study in vitro and in vivo development of Asian yellow goat cloned embryos reconstructed by direct whole cell intracytoplasmic injection (WCICI) into in vitro matured oocytes enucleated at telophase II stage. Our results demonstrated that the rates of cleavage and blastocyst development of embryos reconstructed by WCICI were slightly higher than in conventional subzonal injection (SUZI) group, but no statistic difference (P > 0.05) existed between these two methods. However, the percentage of successful embryonic reconstruction in WCICI group was significantly higher than that in SUZI group (P < 0.05). After embryo transfer at 4‐cell stage, the foster in both groups gave birth to offspring. Therefore, the present study suggests that the telophase ooplasm could properly reprogram the genome of somatic cells, produce Asian yellow goat cloned embryos and viable kids, and whole cell intracytoplasmic injection is an efficient protocol for goat somatic cell nuclear transfer. Mol. Reprod. Dev. 74: 28–34, 2007. © 2006 Wiley‐Liss, Inc.