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KMID : 1005220130280030185
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Journal of Embryo Transfer
2013 Volume.28 No. 3 p.185 ~ p.191
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Development of Cryopreservation Technique of Transgenic Bovine Embryos
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Uhm Sang-Jun
Yang Jung-Seok
Lee Su-Min
Joe So-Young
Lim Joon-Gyo
Heo Young-Tae
Xu Yong-Nan
Koo Bon-Chul
Cheong Ki-Soo
Kim Kwang-Jae
Kim Ji-Tae
Kim Nam-Hyung
Ko Dae-Hwan
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Abstract
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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.
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KEYWORD
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vitrification , PNC , lentivirus , GFP , bovine
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