Journal of Animal Reproduction and Biotechnology 2019; 34(4): 305-310
Published online December 31, 2019
Copyright © The Korean Society of Animal Reproduction and Biotechnology.
Byung-Hyun Choi1,3, Bun-Young Park3, Rami Kong1,3, Mi-Ju Son3, Chan-Sang Park1,3, Nyeon-Hak Shin4, Hye-Young Cheon4, Yeong-Rok Yang4, Jin-Woo Lee4, Jong-In Jin2,3 and Il-Keun Kong1,2,*
1Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju 52828, Korea
2Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 52828, Korea
3GAST, Gyeongsang National University, Jinju 52828, Korea
4Gyeongnam-do Provincial Government Livestock Research Institute, Sancheong 52263, Korea
Embryos produced with serum show the alterations in their ultrastructure, impaired compaction, abnormal blastulation, aberrant mRNA expression profiles and large calf syndrome with greater incidences of stillbirths and deaths after birth. The aim of the present study was to describe
Keywords: embryo development, free-serum, Korean native cow (Hanwoo), OPU
The aim of the present study was to describe
The OPU-IVP data used in this study from 2016. Sixteen Korean native cows (Hanwoo) approximately, which belonged to the Institute of Agriculture and Life Science, Gyeongsang National University, were used. Study based on 2 experimental groups. Donor is equivalent. Serum group for March to July and free-serum group for September to December.
OPU derive oocyte is (TCM-199) supplemented with 10% (v/v) FBS (Gibco BRL, Life Technologies, Grand Island, NY, USA), 1 μg/mL estradiol-17β, 10 μg/mL follicle-stimulating hormone, 10 ng/mL epidermal growth factor, 0.6 mM cysteine, and 0.2 mM sodium pyruvate. Thereafter, groups of up to 50 COCs were transferred to a 4-well dish (Thermo Fisher Scientific, Waltham, MA, USA) containing 500 μL IVM medium and incubated in a humidified atmosphere of 5% CO2 in air at 38.5°C for 22-24 h.
Data are presented as mean values standard deviation, except for data reporting changes during embryo development, which are presented as percentages. Each experiment was performed at least three times, and results were analyzed with SPSS (ver. 18.0, SPSS Inc., Chicago, IL, USA). Differences between means were considered significant at
The recovered cumulus-oocyte complexes were morphologically classified into four grades based on the number of cumulus cells layers and homogeneity of the cytoplasm (Table 1). Oocytes number for session was greater in serum groups than those in free-serum groups (29.61-0.63 vs. 15.6-0.62;
The results regarding the cleaved embryo, as well as the blastocyst development are shown in Table 2. Embryo cleaved competence rate in free-serum groups was higher than those in serum (62.1% vs. 58.3;
The results regarding the embryo transfer, the number of pregnancy, as well as the pregnancy rate are shown in Table 3. Embryo transfer for 986 recipients. Based on high quality embryo, recipients were allocated into two groups: Group 1, embryo production from serum treatment; Group 2, embryo production from free-serum treatment. Pregnancy rate were result of serum was no difference than that in free-serum (54.6% vs. 56.3%;
This study aimed to evaluate the suitability of a serum-free culture system for OPU derived bovine embryos. Blastocyst quality is crucial factor to ensure the optimal pregnancy rates following transfer (Rizos et al., 2003). Serum in culture medium plays a role positive embryotrophic factors and inactivation of emryotoxic agents and lead to improve the embryo development, but their mechanism was not determined (Bavister et al., 1995). The effect of serum on lipid metabolism can be due to three mechanisms: (a) lipoproteins present in the serum can be internalized by the cells, increasing the intracellular lipid content (Sate et al., 1999; Abe and Hoshi, 2003), (b) the presence of serum can alter mitochondria and thus b-oxidation of lipids, leading to increased storage of intracellular lipids (Crosier et al., 2001; Abe et al., 2002) and (c) serum could increase neosynthesis of triglycerides by the embryo (Razek et al., 2000). Hatching rate was able to consider to some extent as a measure of embryo quality. To reproduce the culture conditions usually observed after OPU, we compared the impact of the serum-free culture media and serum media on embryos culture. We investigated the development rate of blastocysts, grade of oocytes, recovery of oocytes and number of follicles according for OPU-derived embryo production. The mean number of oocytes obtained by OPU session varies from 7.2 to 20.9.7 (Galli et al., 2001; Su et al., 2012; Jin et al., 2015; Roth et al., 2018), these data agree with our results reported that the average embryo production per donor were 29.6 and 15.6 embryos at sessions, respectively. However, large variations were observed between breeds and donors. Whatever the culture medium, no difference was observed on embryo development. These results are in agreement with two previous reports (Donnay et al., 1997; Ward et al., 2000). The results of embryo transfer were not different between serum and serum free culture medium. In present results indicate that the free-serum culture media was suitable to use for OPU derived embryo production.
In conclusion, we developed the serum-free system for production of
No potential conflict of interest relevant to this article was reported.
This work was supported by IPET through Agri-Bio Industry Technology Development Program, funded by MAFRA (Grant number: 117029-3 and 315017-5).
BH Choi, Gyeongsang Nat’l Univ., Researcher, https://orcid.org/0000-0001-9768-1962
BY Park, Gyeongsang Nat’l Univ., Researcher, https://orcid.org/0000-0001-7313-7858
R Kong, Gyeongsang Nat’l Univ., Researcher, https://orcid.org/0000-0003-4575-1994
MJ Son, Gyeongsang Nat’l Univ., Researcher, https://orcid.org/0000-0003-1580-6974
CS Park, Gyeongsang Nat’l Univ., Researcher, https://orcid.org/0000-0002-3906-1674
NH Shin, Gyeongnam-do Provincial Government Livestock Research Institute, Ph.D., https://orcid.org/0000-0002-9069-0601
HY Cheon, Gyeongnam-do Provincial Government Livestock Research Institute, Researcher, https://orcid.org/0000-0003-0814-6277
YR Yang, Gyeongnam-do Provincial Government Livestock Research Institute, Researcher, https://orcid.org/0000-0002-7699-0923
JW Lee, Gyeongnam-do Provincial Government Livestock Research Institute, Director, https://orcid.org/0000-0001-9001-7858
JI Jin, Gyeongsang Nat’l Univ., Ph.D., https://orcid.org/0000-0002-5790-7637
IK Kong, Gyeongsang Nat’l Univ., Professor, https://orcid.org/0000-0002-1906-9898