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http://dx.doi.org/10.12750/JET.2013.28.3.185

Development of Cryopreservation Technique of Transgenic Bovine Embryos  

Uhm, Sang Jun (Department of Animal Science and Biotechnology, Sangji Youngseo College)
Yang, Jung Seok (Department of Animal Science and Biotechnology, Sangji Youngseo College)
Lee, Su Min (Department of Animal Science and Biotechnology, Sangji Youngseo College)
Joe, So Young (Department of Animal Science and Biotechnology, Sangji Youngseo College)
Lim, Joon Gyo (Department of Animal Sciences, Chungbuk National University)
Heo, Young-Tae (Department of Animal Sciences, Chungbuk National University)
Xu, Yong-Nan (Department of Animal Sciences, Chungbuk National University)
Koo, Bon-Chul (Department of Physiology, Catholic University of Daegu School of Medicine)
Cheong, Ki-Soo (South Branch of Gangwondo Veterinary Service LAB)
Kim, Kwang Jae (South Branch of Gangwondo Veterinary Service LAB)
Kim, Ji Tae (South Branch of Gangwondo Veterinary Service LAB)
Kim, Nam-Hyung (Department of Animal Sciences, Chungbuk National University)
Ko, Dae Hwan (Department of Animal Science and Biotechnology, Sangji Youngseo College)
Publication Information
Journal of Embryo Transfer / v.28, no.3, 2013 , pp. 185-191 More about this Journal
Abstract
The purpose of this study is to improve production efficiency of vitrified-thawed transgenic bovine embryos. Transgenic bovine embryos were produced by injection of FIV-GFP lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization. EGFP-expressing transgenic bovine blastocysts were cultured in serum-containing and serum-free medium. These blsatocysts were vitrified by pull and cut (PNC) container made with 0.25 cm plastic straw. Results indicate that total developmental rates of normal IVF embryo cultured in serum-containing and-free medium into blastocyst were not significantly different (22.3 vs 21.5%) and those of GFP-expressing transgenic bovine embryo into blastocyst showed no significant difference between serum-containing (13.9%) and-free medium (13.1%). However, developmental rate of GFP transgenic embryo was significantly (P<0.05) lower than its of normal IVF embryos. In additional study, we vitrified GFP transgenic normal bovine blastocysts using PNC vitrification method. Survival rate of vitrified-thawed GFP transgenic blastocyst (23.1%) was significantly (P<0.05) lower than its of normal blastocysts (68.9%). Although, survival rate of vitrified-thawed GFP transgenic blastocyst was lower than its of normal blastocyst, our result may suggested that PNC vitrification method is feasible to cryopreserve transgenic embryos. Our next plan will be the production of GFP express transgenic bovine derived from vitrified-thawed embryos using PNC method.
Keywords
vitrification; PNC; lentivirus; GFP; bovine;
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