This study was carried out to determine the effect of cumulus cell attachment and various factors on in vitro maturation of pig foflicular oocytes. Oocytes with various configuration of cumulus cell mass were collected ftom ovaries of mature gilts by asperating with syringe equipped with needles of different gauges, follicle size and with or without cumulus cells. They were cultured in TCM-199 mediun containing FGS(fetal calf serum) for 30~48 hours in incubator with air containing 5% at 38.5. Mter orcein staining at in vitro maturation condition, GV, GVBD, anaphase, telophase and M II were observed. Results are surumarized as follows: 1. Recovery rates were 55.8, 55.5 and 34.4% when the cumulus-compacted oocytes were collected with 18, 21, 26 gauge needles of syringes, respectively. 2. 79% of oocytes with compacted cumulus cells were at GV stage and most of the oocytes with partially denuded and denuded cumulus cells were from GVBD to M- II stages. 3. Percentage of mature oocytes among those which are follicular diameter of 1~2, 3~6 and over 6 mm was 42.6, 53.2 and 60.8%, respectively. 4. Percentage of mature oocytes among those which are compacted, partially denuded and denuded was 60.5, 46.2 and 35.4% respectively. 5. Percentage of mature oocytes in co-cultured with monolayers of cumulus cells was higher (57.1%) than that found with oocytes cultured alone (53.4%).

The aim of this study was to evaluate the effect of recombinant bovine somatotropin (bST; Boostin-S, LG Chem) treatment with FSH (Super OV) or PMSG on superovulatory response for transvaginal ultrasound-guided oocyte retrieval (TVR) in calves. Eight Korean Native Cattle(KNC) heifer calves; 150 to 240 days old; were randomly assigned to four treatment groups: 1) FSH(75 mg); 2) FSH (75 mg) + bST(500mg) 3) PMSG(1;000 IU); 4) PMSG(1, 000 IU) + bST(500 mg). Experimental calves in group 1 (n=2) and 2(n=2) were weekly superovulated for 4 consecutive weeks with daily injection of FSH for 3days and the next day subjected to TVR session. Animals in group 3 (n=2) and 4(n=2) were weekly stimulated for 4 consecutive weeks with a single dose of 1, 000 IU PMSG. TVR was performed on 72 hours after PMSG injection. Calves in group 2 and 4 was received injection of 500 mg of bST every 10 days. At each TVR session, follicle number and size were recorded; the oocytes collected and graded according to cumulus and cytoplasm investment. Collected oocyte were determined viable oocyte according to morphological quality with granulation of oocyte and number and status of cumlulus cells. IVM and IVF were performed and assessed cleavage rate on day 3 after fertilization. A Sonovet 600(Medison, Co., Ltd) realtime ultrasound scanner with a 6.5 MHz convex transducer, fixed at the tip of 500 mm estended handle equipped with a needle guide was used in collecting oocyte. Differences between groups were analysed by chi-square test. The population of large follicle (5 nun) and aspiration rate were not significant different among the 4 groups. But, the number of small follicles (<5 mm) and aspirated oocyte in the KNC calves treated with bST were 1.3~1.6 times higher than in KNC calves treat with FSH or PMSG alone. In conclusion, the administration of bST with FSH or PMSG at superovulation for TVR in calves was increase the nurnber of small follicle which was influenced the number of aspiratable follicle.

This experiment was carried out to study the determination of survival of vitrified and thawed mammal follicular oocytes by FDA-test. Oocytes were divided into 3 groups according to attachment of cumulus cell. Group A oocytes were tightly surrounded by cumulus cell, group B oocytes were partially surrounded by cumulus cell, and group C oocytes were poorly surrounded by cumulus cell. Vitrification solution developed by our previous study (Kim et al, 1992) which consisted of permeable agent (20 % glycerol + 10 % ethylene glycol) and nonpermeable agent (30 % Ficoll + 10 % sucrose). Oocytes (7~10) loaded into 0.25 ml straw after 10 min equilibration were plunged into liquid nitrogen (- 196) directly. The FDA-score of vitrified and thawed group A oocytes was higher in rat (4.2) than in rabbit (3.9), cow (3.8), mouse (3.4) and porcine (2.4), however that of cumulus cell was higher in rabbit (4.7) than in rat (4.1), cow (2.9), porcine (2.6) and mouse (1.4). The FDA-score of vitrified and thawed group B oocytes were 3.1 (cow), 2.9 (rabbit), 2.9 (mouse), 2.6 (rat) and 2.5 (porcine), respectively. However that of cumulus cell was higher in rabbit (3.7) than in porcine (2.6), rat (2.3), cow (1.7) and mouse (0.3). The FDA-score of vitrified and thawed group C oocytes was higher in mouse (4.1) than in cow (2.9), rabbit (2.6), rat (1.3) and porcine (1.1). As shown in the above results, The survival rates of oocytes were higher in group A than in group B and C except in mouse and cow. These results suggest that the survival of cumulus cell as well as follicular oocytes can be reliably judged by their fluorescence with FDA-test.

This study was carried out to investigate On in vitro fertilization, survival rate and developmental rate of rapidly frozen bovine immature oocytes. Immature oocytes cultured for 1, 12, 24, 48 hours in 20% FCS + TCM-199 medium and thereafter rapidly freezing-thawed oocytes inseminated with capacitated sperm. The immature oocytes following dehydration by 1.5M DMSO + 2.0M glycerol + 0.25M sucrose + TCM 199 media + 20% FGS were directly plunged into liquid nitrogen and thawes in 3 water. Rapid freezing embryos co-cultured in 20% FCS + TCM-199 media containing hormones(21U/mL PMSG, 21U /mL hGG and 1 g /mL 17-estradiol) and cumulus cells(1 x 105-6 cells). Survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro maturation and fertilization rate of immature bovine oocytes on in vitro maturation period(1, 12, 24, 48 hrs) before rapid freezing4hawed were 57.1%, 45.7%, 37.1%, 25.7% and 40.0%, 31.4%, 20.0%, 11.4%, respectively. 2. The survival rate of immature bovine oocytes on in vitro maturation period(1, 12, 24, 48 hrs) before rapid freezing-thawed were 33.3%, 26.7%, 20.0%, and 10.0%, respectively. The survival rate of rapid freezing4hawed immature oocytes was significantly lower than that of non-freezing oocytes. 3. The survival rate of rapid freezing4hawed excellent and good bovine embryos co-cultured in 20% FCS + TCM-199 media containing hormones(PMSG, hCG, 17-estradiol) and cumulus cells 4 to 5 hrs and 20 to 24 hrs were 35.0%, 15.0% and 25.0%, 15.0% and 40.0%, 20.0% and 30.0%, 15.0%, respectively. The survival rate of embryos co-cultured in TCM-199 media containing hormones and cumulus cells was significantly higher than that of non co-culture.

소 난자의 시험관내 수정시 이성체인 1, 2 propanediol과 1, 3 propanediol과의 동결보호제의 이용에 대한 독성효과를 조사하였다. 프랑스에 있는 도축장의 난소에서 채취한 미성숙 난자 212개를 시험관내 성숙과 heparin(10g/ml)첨가에 의한 수정 및 배양을 실시하였다. 소 난자의 시험관내 성숙과 수정시 propanediol의 독성효과를 시험한 결과 1,3 propanediol은 80%의 정상 난할분할율과 5.7%의 낮은 다핵분

The present work was designed to understand the mechanism of superovuiation and the cause of early embryo loss and Implantation failure in the superovulating immature female rats which were elaborated by a pituitary gland transplantation. A pituitary gland obtained from the orchidectomized rats was transplanted under the right kidney capsule of 28 day old female rats (PGT group) on the starting day of experiment which was designated as Day 2. The grafted pituitary glands were removed at 6h (RPGT 6h group), 12h (RPGT 12hgroup) and 24h (RPGT 24h group) after the transplantation. Control rats were treated with 41U PMSG on Day 2 (PMSG group). The estrous cycle and the levels of plasma progesterone and estradiol-17 were observed on Day 0, Day 5, respectively. The implantation sites, the weights of ovary and uterus, and the number of corpora lutea were examined in all group on Day 8. The resuft obtained were summarized as follows: 1. The percentages of the number of the rats in proestrus and estrus were 93.0%, 82.6%, 0%, 90.7% and 89.5% in PMSG, PGT, RPGT6h, RPGT12h and RPGT24h group, respectively. The synchronization of estrus cycle was dchieved in all groups. 2. The mating rates of each group were 80.2, 75.0, 0, 56.4, 57.8% in PMSG, PGT, RPGT6h, RPGT12h, RPGT24h group, respectively. 3. The numbers of copora lutea on Day 8 were 47.1 i 4.9, 18.1 0.5, 14.1 i 0.3 and 8.9 0.3 in PGT, RPGT24h, RPGTl2h and PMSG group, respectively. There were signIficantly difference between all groups (P<0.05). 4. The numbers of implantation sites (18.1 +- 4.0) in PGT group on Day 8 were higher than those of PMSG (8.5 2.5), RPGT 12h (9.8 i 0.2) and RPGT 24h group (10.8 i 0.2) (P<0.05). 5. The ovarlan weights in PGT (95.2 14.3mgIlOOg BW), RPGT 12h (51.7 0.6mgIlOOg BW), and RPGT24h (57.9 0.9mg/l00g 8W) groups were significantly higher than those of PMSG group (30.4 7.4mg/l00g BW) (P<0.05). 6. The uterine weights in PMSG (672.4 4.7mg/l00g 8W), and PGT (660.7 7.8mg/l00g BW) groupswere greater than those of RPGT 12h (403.0 1.lmg/lOOg BW) and RPGT 24h (490.1 0.9mg/l00g BW) group (P<0.05). 7. The plasma progesterone levels in PGT groups (15 lng/ml) on Day 5 were higher than those of PMSG (83ng/ml), RPGT 12h (S7ng/ml), RPGT24h (8lng/ml) group (P<0.05). 8. The plasma estradiol-17 levels in PMSG group (lBSpg/ml) on Day S were higher than those of RPGT 24h (l3pg/ml) group (P<0.05). But estradiol-17 levels in PGT and RPGT 12h group were too low to discuss.

부신(副腎)의 기능(機能)이 생식기관(生殖器官)에 미치는 영향(影響)을 구명(究明)코자 21일령(日齡)의 암흰쥐(體重(체중): ) 210마리를 정상(正常)흰쥐실험군(實驗群)(이하(以下) Int.-)과 부신척출실험군(副腎剔出實驗群)(이하(以下) Adx.-)으로 나누고, 각군(各群)을 다시 대조군(對照群)(이하(以下)-Cont.), PMS 투여군(投與群)(이하(以下) - PMS) 및 PMS와 cortisol의 병용투여군(이하(以下)-PMS+corti)의 3개군(個群)으로 나누어 35마리씩 배치(配置)한 다음 부신척출실험군(副腎剔出實驗群)에 대(對)하여 부신척출(副腎剔出)을 실시(實施)한 5일후(日後)(흰쥐 일령(日齡) : 26일(日))에 PMS투여군(投與群)에는 마리당 25IU의 PMS를, PMS와 Cortisol의 병용투여군에는 마리당 25IU의 PMS와 의 Cortisol의 처리후(處理後) 48, 54, 60, 66, 72, 78 및 84시간(時間)에 5마리씩 도살(屠殺)하여 생식기능(生殖機能)의 중량(重量)을 측정(測定)하였다. 1. 난소(卵巢)의 중량(重量)은 모든 처리군(處理群)이 처리후(處理後) 기간(期間)이 경과(經過)할수록 증가(增加)하여 전관찰시간(全觀察時間)에서 비교군간(比較群間)에 고도(高度)의 유의성(有意性)(P<0.01)이 인정(認定)되었으나 대조군(對照群)인 Int.-Cont. 군(群)과 Adx.-Cont. 군(群)은 변화(變化)가 인정(認定)되지 않았다. 다중검정결과(多重檢定結果) 부신(副腎)을 척출(剔出)한 Adx.-PMS군(群)과 Adx.-PMS+Corti. 군(群)은 정상(正常)흰쥐 실험군(實驗群)인 Int.-PMS군(群)과 Int.-PMS+Corti. 군(群)에 비하여 유의(有意)(P<0.05)하게 낮은 값을 보였고, Cortisol 투여(投與)에 따른 효과(效果)는 인정(認定)되지 않았다. 2. 자궁(子宮)의 중량(重量)은 모든 관찰시간(觀察時間)에서 비교군간(比較群間)에 고도(高度)의 유의성(有意性)(P<0.01)이 인정(認定)되었는데, 처리후(處理後) 66시간(時間)까지는 정상(正常)흰쥐 실험군(實驗群)인 Int.-PMS군(群)과 Int.-PMS+Corti.군(群)이 높은 값을 보였고, 72시간(時間) 후(後)부터는 반대(反對)로 부신척출실험군(副腎剔出實驗群)인 Adx.-PMS군(群)과 Adx.-PMS+Corti.군(群)이 높은 값을 나타냈으며, 다중검정결과(多重檢定結果) Int.-PMS군(群)과 Int.-PMS-Corti.군(群), 그리고 Adx.-PMS군(群)과 Adx.-PMS군(群)과 Adx.-PMS+Corti.군(群) 상호간(相互間)에는 유의성(有意性)이 인정(認定)되지 않았다. 3. 정상(正常)흰쥐 실험군(實驗群)에서의 부신(副腎)의 중량(重量)은 Int.-PMS 군(群)에서 근소(僅少)하게 높은 값을 나타냈으나 비교군간(比較群間)에는 통계적(統計的)인 유의성(有意性)은 인정(認定)되지 않았고, 조직소견(組織所見)에서도 특기(特記)할 만한 변화(變化)를 인지(認知)할 수 없었다.