Vitamin C enhances porcine cloned embryo development and improves the derivation of embryonic stem-like cells |
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| Affiliation: | 1. Laboratory of Theriogenology, College of Veterinary Medicine, Chungnam National University, Daejeon, Republic of Korea;2. Research Institute of Veterinary Medicine, Chungnam National University, Daejeon, Republic of Korea;3. Department of Transgenic Animal Research, Optipharm, Inc., Chungcheongbuk-do, Cheongju-si, Republic of Korea;1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, 730070, Gansu, China;2. Gansu Key Laboratory of Animal Generational Physiology and Reproductive Reg-ulation, China;1. Department of Gynecology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, China;2. Department of Dermatology, Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine, China;3. Department of Medical Imaging, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, China;1. Department of Science and Education, The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, Guangxi, PR China;2. The Reproductive Medical Center, The Second Nanning People’s Hospital, Nanning, 530031, Guangxi, PR China;3. Department of Reproductive Medicine, The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, Guangxi, PR China;4. Department of Clinical Laboratory, The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, Guangxi, PR China;5. Guangxi University of Chinese Medicine, Nanning, 530200, Guangxi, PR China;1. Laboratorio de Medicina Reproductiva, Universidad Nacional de Rosario, Suipacha 531, C.P: 2000, Rosario, Santa Fe, Argentina;2. Consejo de Investigaciones de la Universidad Nacional de Rosario (CIUNR), Rosario, Santa Fe, Argentina;3. Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET, Rosario, Santa Fe, Argentina;1. Clinic of Reproductive Medicine, Vetsuisse Faculty, University of Zurich, Switzerland;2. Department of Theriogenology, Faculty of Veterinary Medicine, Cairo University, Egypt;3. Gestüt Lewitz, Neustadt-Glewe, Germany;4. Institute of Veterinary Anatomy, Vetsuisse Faculty, University of Zurich, Switzerland |
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| Abstract: | Porcine cloning through somatic cell nuclear transfer (SCNT) has been widely used in biotechnology for generating animal disease models and genetically modified animals for xenotransplantation. Vitamin C is a multifunctional factor that reacts with several enzymes. In this study, we used porcine oocytes to investigate the effects of different concentrations of vitamin C on in vitro maturation (IVM), in vitro culture (IVC), and the derivation of nuclear transfer embryonic stem-like cells (NT-ESCs). We demonstrated that vitamin C promoted the cleavage and blastocyst rate of genetically modified cloned porcine embryos and improved the derivation of NT-ESCs. Vitamin C integrated into IVM and IVC enhanced cleavage and blastocyst formation (P < 0.05) in SCNT embryos. Glutathione level was increased, and reactive oxygen species levels were decreased (P < 0.05) due to vitamin C treatment. Vitamin C decreased the gene expression of apoptosis (BAX) and increased the expression of genes associated with nuclear reprogramming (NANOG, POU5F1, SOX2, c-Myc, Klf4, and TEAD4), antioxidation (SOD1), anti-apoptotic (Bcl2), and trophectoderm (CDX2). Moreover, vitamin C improved the attachment, derivation, and passaging of NT-ESCs, while the control group showed no outgrowths beyond the primary culture. In conclusion, supplementation of vitamin C at a dose of 50 µg/ml to the IVM and IVC culture media was appropriate to improve the outcomes of porcine IVM and IVC and for the derivation of NT-ESCs as a model to study the pre- and post-implantation embryonic development in cloned transgenic embryos. Therefore, we recommend the inclusion of vitamin C as a supplementary factor to IVM and IVC to improve porcine in vitro embryonic development. |
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| Keywords: | Cloned embryo Transgenic animals Embryonic stem-like cells Vitamin C Oocyte maturation |
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