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Effect of $\beta$-Mercaptoethanol on In Vitro Maturation of Porcine Folliculay Oocytes and Development of Porcine IVM/IVF Embryos  

한만희 (축산기술연구소)
이경본 (충남대학교 동물자원학부)
천행수 (충남대학교 동물자원학부)
박병권 (공주대학교 영상보건대학 특수동물학과)
서길웅 (충남대학교 동물자원학부)
이규승 (충남대학교 동물자원학부)
Publication Information
Abstract
The present study was carried out to examine the effect of $\beta$-Mercaptoethanol ($\beta$-ME) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with $\beta$-ME on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows. 1. The rates of nuclear maturation, penetrated oocytes, polyspermic oocytes, pronucleus formation and mean numbers of the penetrated sperms were not significantly different using NCSU-23 maturation media for 0, 25, 50 and 100 $\mu$M $\beta$-ME (P>0.05). 2. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in oocytes matured with 25 $\mu$M $\beta$-ME (25.4$\pm$0.9%) than in those matured with 0 (14.5$\pm$1.6%), 50 (17.3$\pm$1.7%) and 100 $\mu$M (12.4$\pm$1.3%) (P<0.05). However, no differences ware found in total cell numbers of blastocyst among the treatments. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in the NCSU-23 Culture medium With 25 $\mu$M $\beta$-ME (23.6$\pm$2.8%) than in those Cultured With 0 (15.4$\pm$4.4%), 12.5 (17.5$\pm$2.3%) and 50 $\mu$M $\beta$-ME (18.6$\pm$2.1%) Under the 5% and 20% $O_2$ Concentrations (P<0.05). However, no differences was found in total cell numbers of blastocyst among the treatments. These results suggested that the addition of 25 $\mu$M $\beta$-ME in the IVM/IVD media were effective on the porcine embryo production. However, the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not significantly different in the NCSU-23 culture medium under 5% and 20% 02 concentrations.
Keywords
Porcine embryo; IVM/IVF; $\beta$-ME; $\beta$-mercaptoethanol;
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1 Byun, T. H., Lee, S. H. and Song, H. B. 1991. Development of a rapid staining method of the oocytes from domestics animal. Kor. J. Anim. Sci. 33:25-31
2 Edwards, R. G. 1965. Maturation of mouse, sheep, cow, pig, rhesus monkey, and human ovarian oocytes. Nature. 208:349-352   DOI   PUBMED   ScienceOn
3 Lim, J. M., Reggio, B. C., Godke, R. A. and Hansel, W. 1999. Development of in-vitro derived bovine embryos cultured in 5% $CO_2$ in air or in 5% $O_2$, 5% $CO_2$ and 90% $N_2$ Human Reprod. 14(2):458-464
4 Lim, J. M., Liou, S. S. and Hansel, W. 1996. Intracytoplasmic glutathione concentration and the role of $\beta$-mercaptoethanol in preimplantation development of bovine embryos. Theriogenology. 46:429-439.   DOI   ScienceOn
5 이홍준, 서승훈, 이광희, 김기동, 이상호, 송해범. 1997. $\beta$-Mercaptoethanol 첨가에 의한 소 초기배의 체외발생 효과. 한국가축번식학회지. 21(4):389-396
6 Mattioli, M., Bacci, M. L., Galeati, G. and Seren, E. 1989. Developmental competence of pig oocytes matured and fertilized in vitro. Theriogenology. 31:1201-1209
7 Park, C. K., Roy, F. and Sirard, M. A. 1996. The effect of free radicals and antioxidant during in vitro maturation and fertilization of porcine oocytes. Theriogenology. 45:275(abstr.)
8 Geshi, M., Yonai, M., Sakaguchi, M. and Nagai, T. 1999. Improvement of in vitro co-culture systems for bovine embryos using a low concentration of carbon dioxide and medium supplemented with p-mercaptoethanol. Theriogenology. 51:551-558   DOI   ScienceOn
9 Iritani, A., Niwa, K. and Imai, H. 1978. Sperm penetration in vitro of pig follicular oocytes matured in culture. J. Reprod. Fert. 54:379-383   DOI   ScienceOn
10 Abeydeera, L. R. and Day, B. N. 1997. In vitro penetration of pig oocytes in a modified trisbuffered medium: effect of BSA, caffeine and calcium. Theriogenology. 48:537-544   DOI   ScienceOn
11 Takahashi, M., Nagai, T., Hamano, S., Kuwayama, M., Okamura, N. and Okano, A. 1993. Effect of thiol compounds on in vitro develpoment and intracellular glutathione content of bovine embryos. BioI. Reprod. 49: 228-232
12 Machaty, Z., Day, B. N. and Prather, R. S. 1998. Development of early porcine embryos in vitro and in vivo. BioI. Reprod. 59:451-455   DOI   ScienceOn
13 Caamano, J. N., Pugh, M. L., Rowson, A. D. and Youngs, C. R. 1998. Effect of oxygen tension and $\beta$-mercaptoethanol on development of in vitro produced bovine embryos. Theriogenology. 49:196(abstr.)
14 Ishii, T., Bannai, S. and Sugita, Y. 1981. Mechanism of growth stimulation of L1210 cells by 2-mercaptoethanol in vitro: role of the mixed disulfide of 2-mercaptoethanol and cysteine. J. BioI. Chem. 256(23):12387-12392
15 Sagara, J., Miura, K. and Bannai. 1993. Cysteine uptake and glutathione level in fetal brain cells in primary culture and in suspension. J. Neurochem. 61:1667-1671
16 Petters, R. M. and Wells, K. D. 1993. Culture of pig embryos. J. Reprod. Fert.(suppl.) 48: 61-73
17 SAS Insitute. 1996. SAS/STAT$^{\circledR}$ user's guide, release 6.12 edition. SAS Institute Inc., Cary, NC. USA
18 Caamano, J. N., Ryoo, Z. Y., Thomas, J. A. and Youngs, C. R. 1996. $\beta$-Mercaptoethanol enhances blastocyst formation rate of bovine in vitro-matured/in vitro-fertilized embryos. BioI. Reprod.55:1179-1184   DOI   ScienceOn
19 양부근, 박동헌, 정희태, 박춘근, 김종복, 김정익. 1997. Thiol 화합물과 항산화제 첨가배양이 소 체외수정란의 체외발육과 세포내 Glutathione 농도 변화에 미치는 효과: I .$\beta$-Mercaptoethanol과 Cysteamine 첨가가 소 체외수정란의 체외발육과 세포내 Glutathione 농도변화에 미치는 영향. 한국가축번식학회지. 21(4): 335-343
20 Abeydeera, L. R., Wang, W. H., Cantley, T. C., Prather, R. S. and Day, B. N. 1998. Presence of $\beta$-mercaptoethanol can in increase the glutathione content of pig oocytes matured in vitro and the rate of blastocysts development after in vitro fertilization. Theriogenology. 50:747-756   DOI   ScienceOn
21 Cheng, W. T. K., Polge, C. and Moor, R. M. 1986. In vitro fertilization of pig and sheep oocytes. Theriogenology. 25:146(abstr.)
22 Hunter, R. H. and Polge, C. 1966. Maturation of follicular oocytes in the pig after injection of human chorionic gonadotrophin. J. Reprod. Fert. 12:525-531   DOI   ScienceOn