Korean Journal of Veterinary Research 1989;29(2):123-128.
Study on improvement of viability of mouse embryos after bisection
Hyo-jong Lee1, Hee-sung Park2, Taeg-seog Kim1, Sang-yong Choe1, Choong-saeng Park2
1College of Veterinary Medicine, Gyeongsang National University
2College of Agriculture, Gyeongsang National University
생쥐 수정란의 분할조작후 생존성 향상에 관한 연구
이효종1, 박희성2, 김택석1, 최상용1, 박충생2
1경상대학교 수의과대학
2경상대학교 농과대학
Abstract
Demi-embryos were successfully produced by bisection of ICR mouse embryos at preimplantation stages. They were microsurgically bisected using a microsurgical blade attached to a micromanipulator after pretreatment with 0.5% pronase in PBS for two minutes or not. Embryos with softened zona pellucida were more easily bisected and less damaged than intact embryos. The highest success rate in bisection has been achieved by selecting blastocysts(94.1% in success rate with intact blastocysts and 100% in success rate with zona softened blastocysts). Demi-embryos without zona pellucida were cultured in D-PBS or M-16 medium at $37^{circ}C$, 5% $CO_2$ in air for 72 hours for 2-cell stage embryos, 48 hours for 4-to 8-cell stage embryos, 24 hours for morula stage embryos and 6~12 hours for blastocyst stage embryos. For the in vitro culture of 2-cell stage embryos, $100{mu}M$ 2Na-EDTA was added to the media. M-16 medium was better for the in vitro development of mouse embryos than PBS, and PBS is not considered to be suitable for long-term culture of embryos, especially at early stage of cleavage. In M-16 medium, developing rate of demi-embryos of which pair underwent development to form eublastocysts was 15.8% at 2-cell stage, 16.8% at 4-cell stage, 38% at 8-cell stage, 89.6% at morula stage and 94.4% at blastocyst stage, respectively. The more rapid and efficient production of demi-embryos and higher viability after bisection can be expected by softening zona pellucida with pronase and by selecting morulae or blastocysts rather than embryos at early stage of cleavage.
Key Words: embryo bisection, viability, mouse


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