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KMID : 1005220140290010073
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Journal of Embryo Transfer
2014 Volume.29 No. 1 p.73 ~ p.81
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Effects of Vitamin K1 on the Developmental and Survival Rate of Porcine In Vitro Fertilized Embryos
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Park Hum-Dai
Zhu Yi-Chen
Park Yong-Soo
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Abstract
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The in vitro production of porcine embryos was essential to increase of blastocyst development rate and select of high quality blastocyst in early stage. There were a lot of reports about in vitro porcine embryo development, but there was no report about the selection of high quality embryos. Therefore, in this study, we investigated the effect of vitamin K1 (vit K1) on the development and survival rate of porcine in vitro fertilized embryos. When vit K1 was treated for 24 hr at day 1 in vitro culture, blastocyst development rate in the control group (35.5 ± 3.2%) was significantly lower compared to 1.0 μM, 3.0 μM, or 6.0 μM groups (14.5 ± 4.3, 0.0, or 0.0%; p<0.05). The survival rates of blastocysts at day 8 in 1.0 μM, 3.0 μM or 6.0 μM of vit K1 treated groups (22.2 ± 2.9, 0.0 or 0.0%) were significantly lower than that of the control group (31.8 ± 2.6%; p<0.05). We were added at 1.0 μM, 3.0 μM or 6.0 μM vit K1 for different durations of time at day 1 in vitro culture. The development rate and survival rate in the group of 1.0 μM vit K1 for 6 hr was 26.5 ± 2.9% and 47.2 ± 2.8%, respectively, which were differed significantly in the group of 12 hr (p<0.05). In the group of 3.0 μM vit K1, the blastocyst development in control group was 36.4 ± 3.1% but, the survival rate 41.7 ± 3.2% in the group of 3.0 hr was significantly higher than that of the control group (p<0.05). In the group of 6.0 μM vit K1, the control group’s the blastocyst development was 32.0 ± 2.8% and the 0.5 hr supplement group’s survival rates was 42.9 ± 1.8% higher than other groups. We added vit K1 at day 1, day 2, day 4 and day 6 of in vitro culture, on the based the results of supplemented concentration and duration. In the group of 1.0 μM 6.0 hr addition, the blastocyst development rate of day 4 and the survival rate of day 2 were the highest in each group. In the groups of 3.0 μM 3.0 hr addition or 6.0 μM 0.5 hr addition, the blastocyst development (59.5 ± 4.1% and 50.0 ± 3.6%) and survival rates (72.7 ± 5.4% and 79.2 ± 4.0%) on day 4 were significantly higher than that of control and other experiment groups (p<0.05). Meanwhile, the number of cells in blastocysts that produced by vit K1 supplementation was 53.4 ± 5.8, 49.4 ± 3.8 and 51.5 ± 4.5 respectively, which were significantly higher than that of 40.2 ± 2.3 in the control group (p<0.05). There was no difference of the number of apoptotic cells between control and experiment groups. In addition, gene expression of survival blastocyst, the Bax mRNA expression was similar between the control and the experiment groups. However, Bcl-xL mRNA expression’s in the group of 6.0 μM 0.5 hr on day 4 was highest among control and experiment groups (p<0.05). In this study suggested that the control of concentration, duration and time was effective on the survival and cell number of porcine blastocyst derived from in vitro. We are not know what the exact reasons of the effect of vit K1 on embryo development and need to fur ther study. However, vit K1 might be using the selection of high quality porcine blastocyst.
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KEYWORD
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vitamin K1, porcine, in vitro culture, survived blastocyst
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