The aim of this study is to elucidate sperm chemotaxis and to set up the optirnal condition for selection of motile and capacitated sperm from hovine frozen-semen. Thus, the effects of semen-washing after thawing, concentrations of progesterone (P4) and bovine serum albumin (BSA), and sperm-washing frequency on sperm selection were examined. For evaluating their effects, number, viability and acrosome reaction of sperm swim-up seperated from semen, which were incubated for 30 minutes at 36 in the M2 solution containing P4 and BSA, were investigated. For frozen-semen just after thawing, sperm recovery and viability were not significantly different between P4-treated and -untreated semen. However, washing frozen-semen decreased the number of sperm and increased the viability of sperm that were recovered from semen treated with P4. Progesterone affected the recovery rate, the viability and the acrosome-reaction rate of sperm recovered from washed frozen-semen. Especially, number of motile and capacitated sperm were highest in semen treated with 50g /ml among 0, 20, 50 and 100g /ml of P4 concentrations. BSA affected the recovery rate and the viability of sperm recovered from washed frozen-semen that were treated with 50g /ml of P4. Especially, the percentage of viable sperm were highest in semen treated with 4mg /ml among 0, 2, 4, and 6mg /ml of BSA concentrations. Repeatedly sperm-washing did not affect the recovery rate and the viability of sperm recovered from washed frozen-semen that were treated with 50g /ml of P4 and 4mg /ml of BSA In conclusion, using progesterone and BSA could efficiently make the selection of motile and capacitated sperm from washed frozen-semen.

The efficient cryopreservation of embryos requires optimal times of dehydration and rehydration This study was carried out to investigate the effect of various times of dehydration and rehydration The effects were evaluated through testing morphological normality and developmental ability of 1 cell mouse embryos which were ultrarapidly frozen and thawed. The 1 cell embryos were dehydrated for 1.5, 3, 5, and 10 minutes using mPBS-BSA containing 3.SM DMSO and 0.25M sucrose on cooling chamber or on ice. After ultrarapidly frozen and thawed, they were rehydrated for 0, 0.5 and 5 minutes with mPBS-BSA containing 0.25M sucrose at room temperature. The results obtained were as follows: The embryos that were rehydrated for 0.5 minutes showed higher normality than the embryos for 0 and 5 minutes did. The embryos that were dehydrated for 10 minutes showed higher normality than the embryos for 1.5, 3, and 5 minutes did. The developmental ability of normal thawed-embryos was high in 10 minute dehydration treatment compared to other treatments. However, it was not affected by cooling methods (on ice and on cooling chamber) for embryo dehydration.

This study was carried out to efficiently use the ultrarapid freezing method in the cryopreservation of mouse ova. For this, the effects of dehydration method, oval vigour and controlling method on post-thawing viability were investigated. Fresh mouse ova were dehydrated in mPBS with 3.5M DMSO and /or 0.25M sucrose, and directly immersed in L for ultrarapidly freezing. The frozen ova were thawed at 37, rehydrated in mPBS with 0.25M sucrose, and then repeatedly washed in HAM's Fl0 before evaluating the morphological normality of frozen-thawed ova. The results obtained showed that there was difference between treatments in a experiment. 1) The post-thawing viability of ova dehydrated in multi-step (48.413.8%) was higher than that of ova in two-step (40.914.0%). 2) The post-thawing viability of fertilized ova (8714.0%) was significantly(p<0.0l) higher than that of unfertilized ova (5.45.4%). 3) The post-thawing viability of ova dehydrated and rehydrated using a cooling machine (95.84.2%) was significantly(p<0.05) higher than that on ice(84.19.9). In conclusion, in order to efficiently cryopreserve ova in vitro with ultrarapidly freezing method, highly viable embryos should be selected, heavy osmotic shock to the dehydrating ova should be avoided, and embryos in high osmotic condition were dehydrated and rehydrated in a constantly low temperature.

본 연구에서는 C++언어를 이용한 객체지향 프로그래밍 기법으로 매트릭스 연산과 평면뼈대 구조물의 해석이 가능한 PC용 구조해석 프로그램을 개발하였다. 객체지향 프로그래밍에서의 주요 개념인 객체, 클래스, 처리방식, 상속성 및 다형성을 도식화하여 설명하였으며, 매트릭스 연산과 평면뼈대 구조해석에 대한 예제 해석 결과는 이 프로그램의 효율성과 타당성을 보여 주었다. 따라서 본 연구는 객체지향 프로그래밍기법의 특징인 프로그램의 확장성과 재사용성 및 다양한 GUI의 구현가능성을 이용하여 앞으로의 객체지향 유한요소 프로그램 개발에 활용될 것이다.

This study was aimed to develope the new method for bovine sperm capacitation through testing and combinating factors relating to sperm capacitation. For verifying the efficiency of this method on inducing capacitation, in vitro fertilization was carried out. The results obtained were as follows: When pHs of HBS /PR for sperm-washing were 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45 and 8.83, variances between light absorhance differences obtained from sperm pre-washing solution and post-washing solution(VADs) were 0.000, 0.001, -0.001, -0.005, -0.005, -0.021,-0.017,-0.016,and-0.036, respectively. There were significant decreases of VADs in pH 7.69 and pH 8.83. When sperm were firstly, secondly, and thirdly washed with HBS /PR, VADs were -0. 024, - 0.006, and - 0.004, respectively. There was significant decrease of VAD in 1st sperm-washing. When washed-sperm were secondly washed with HBS /PR supplemented with no (con-trol), heparin, CSA, PC12, and BSA, VADs were 0.009, -0.024, -0.008, -0.009, and 0.014, respectively. When the sperm were thirdly washed with HBS /PR with no(control), heparin, CSA, PC12 and BSA, VADs were 0.020, -0.007, 0.005, 0.006, and 0.019, respectively. Only heparin treatment showed the negative values of VADs in 2nd and 3rd sperm-washing. Of oocytes cultured with sperm which were repeatedly washed with heparin and high pH, 52% (57/110) were cleaved over 2 cell stage. However, percent of oocytes parthenogenetically divided was 5%(2 /42).