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Improved Developmental Potential of Transgenic Bovine Embryos Using EGF, IGF and Flavonoid

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한국동물생명공학회(구 한국수정란이식학회) (Journal of Animal Reproduction & Biotechnology)
초록

It is well known that the production of transgenic bovine embryos is more difficult than the production of non-transgenic bovine embryos. We performed whether the quality of transgenic bovine embryos expressing the enhanced green fluorescent protein (EGFP) can be improved when cultured for 4 days from day 4 until day 8 after activation with epidermal growth factor (EGF), insulin-like growth factor (IGF), and flavonoid (F) supplements. The EGFP gene was introduced into bovine IVF embryos using microinjector. In experiment, transgenic bovine embryos were cultured in modified CR1aa medium containing 10% FBS at 38.8℃ in an incubator (5% CO2, 5% O2, and 90% N2) for 8 days and embryos were equally divided into four groups [non-treated group (control), 1 μM IGF+EGF (IE), 10 μM F and 1 μM IGF+EGF+10 μ M F (IEF)] at day 4 embryos. In the result, development rate of F treatment group (41.8%) was higher than that of control (33.3%), IE (23.9%) and IEF groups (28.0%). However, hatching rate was significantly high in IE (53.0%) and IEF (65.0%) groups than in control (42.9%) and F group (42.8%) (p<0.05). The EGFP expression rate was not different among all groups (30.0~33.3%) at blastocyst stage. In comparison of total cell number, IE group (145.2±10.4) was significantly higher than control group (101.4±14.3). Apoptotic index of IE group (1.9%) was the lowest compared with that of control (3.3%), F (3.5%) and IEF groups (4.6%). This result demonstrate that the combination of IGF, EGF and flavonoid can be helpful to improve the development potential and the quality of transgenic bovine embryos.

저자
  • Min-Jee Park(Mirae Cell Bio, Jeju National University Stem Cell Research Center, Jeju National University)
  • Eun-Young Kim(Mirae Cell Bio, Jeju National University Stem Cell Research Center, Jeju National University)
  • Se-Pill Park(Mirae Cell Bio, Jeju National University Stem Cell Research Center, Jeju National University)