• 한국가축번식학회지
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• 제21권3호
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• pp.281-285
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• 1997

The study was conducted to investigate on in vitro developmental rates of bisected bovine embryos co-culture in 10% FCS＋TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 7 days after bisection. In vitro developmental rates was defined as development rates on in vitro culture or FDA-test. The results are summarized as follows : 1. In vitro developmental rates of bisected bovine embryos co-cultured in 10% FCS＋TCM-199 media containing PMSG＋hCG, PMSG＋$\beta$-estradiol, hCG＋$\beta$-estradiol, PMSG, hCG 0 to 3 days and 4 to 7 days were 16.7~30.0% and 11.1~25.0%, respectively. In vitro developmental rates of bisected embryos co-cultured in 10% FCS＋TCM-199 media containing hormones significantly higher than that of non co-culture. 2. In vitro developmental rates of bisected bovine embryos co-cultured 10% FCS＋TCM-199 media containing oviductal epithelial cells 0 to 3 days and 4 to 7 days were 25.0% and 22.2%, respectively.

• 한국수산과학회지
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• 제40권6호
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• pp.374-379
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• 2007

Two experiments were designed to examine short-term effects of human chorionic gonadotropin (hCG), and long-term effects of gonadotropin-releasing hormone agonist (GnRHa), $17{\alpha}-hydroxyprogesterone$ (17P), and $17{\alpha},20{\beta}-dihydroxy-4-pregnen-3-one\;(17,20{\beta}P)$, alone or in combination, on milt production of the starry flounder Platichthys stellatus. In the first experiment, fish were injected with either 200 IU hCG/kg body weight or the same volume of marine fish Ringer's solution (MFRS). In the second experiment, each fish was implanted with a blank cholesterol pellet (control), $200\;{\mu}g$ GnRHa, $500\;{\mu}g$ 17P, or $100\;{\mu}g\;17,20{\beta}P/kg$ body weight alone or in combination. In the first experiment, hCG injection resulted in an increase in the expressible milt volume and a decrease in the spermatocrit (Sct). After pellet implantation in the second experiment, the milt volume was increased in males treated with GnRHa, GnRHa+17P, or $GnRHa+17,20{\beta}P$. On day 7 after hormone pellet implantation, the milt volume began to increase, and on day 14, the milt volume in the $GnRHa+500\;{\mu}g$ 17P group was significantly higher than that in the control group. Compared with the control group, the hormone pellet-treated groups had a significant reduction in the mean Sct and sperm concentration (Sc) at day 7 after pellet implantation, while there were no differences in total sperm number. The results suggest that increases in milt volume are generally associated with decreases in Sct and SC, suggesting that the main mechanism for the increase in milt volume was milt hydration.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.377-381
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• 1999

Objectives: Recent studies, including our own, demonstrated that the novel expression of LH gene in rat gonads and uterus, indicating that the local production and action of the LH-like molecule. In the present study, we investigated whether human uterus also expresses the LH gene. Design: Reverse transcription-polymerase chain reaction (RT-PCR) amplified the cDNA fragments coding $LH_{\beta}$ polypeptide from human endometrium but not from myometrium. Presence of the transcripts for the ${\alpha}$-subunit in human endometrium was also confirmed by RT-PCR. Results: Transcripts for $LH_{\beta}$ subunit were detected in endometrial samples from women with endometriosis. The gene for LH/hCG receptor was expressed in both endometrium and myometrium, showing good agreement with previous studies. Increased level of $LH_{\beta}$ transcript was determined in the endometrium from follicular phase compared to that from luteal phase. Conclusion: Taken together, our findings demonstrated that 1) the genes for LH subunits and LH/hCG receptor are expressed in human uterus, 2) the uterine LH expression was changed during menstrual cycle, suggesting that the uterine LH may playa local role in the control of uterine physiology and function(s).

• 한국수정란이식학회지
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• 제12권1호
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• pp.27-36
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• 1997

The studies were carried out to investigate the effects of co-culture with cumulus cells and oviduct epithelial cells on the in vitro fertilization and cleavage rate of bovine follicular cocytes and to determine the optimum thawing temperature and equilibration time on in vitro developmental rate of frozen bovine embryos. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes were cultured in TGM-199 medium containing 10 IU /ml의 PM SG, 10 IU /ml의 hCG, ip g/ml의 $\beta$-estradiol and 10% FCS for 24~48 hrs in incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. The bovine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquld nitrogen and thawed in 3$0^{\circ}C$ water. Survival rate was defined as developmental rate on in vitro culture or FDA-test. The results are sunanarized as followes :1. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with cumulus cells in TCM499 medium were 75.0~76.8% and 17.3~27.6%, respect-ively. And in-vitro fertilization rates of cumulus-enclosed oocytes(55.4%)were significantly(p<0.05) higher than cumulus-denuded oocytes (23.1%). 2. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with l$\times$ l04cells /ml, 1 x l06cells /ml, lx l08cells /ml and 1 x l015cells /ml oviduct epithelial cells in TCM-199 medium were 74.5~77.8% and 15.7~21.20 respectively.3. The in-vitro fertilization and in vitro developmental rates of bovine oocytes cocultured in '1CM-199 media containing PMSG, hCG, PMSG+hCG. PMSG+$\beta$-estradiol, hCG+$\beta$-estradiol 0 to 40 hrs after insemination were 74.0~77.4% and l8.9~23.l%, re-spectiv ely.4.The survival rates of bovine embryos thawed after rapid freezing in the freezing medium containing a various concentration of sucrose added 1.5M and 2.OM glycerol,DMSO and propanediol were 23.5~31.4% and 20.6~34.l%, respectively. 5. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher embryos survival rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$.6. The equilibration time on the survival rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20min.). (Key words : bovine embryos, co-culture, freezing, in vitro development)

• 한국가축번식학회지
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• 제19권2호
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• pp.135-140
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• 1995

The study was conducted to investigate on the survival rate of bisected porcine embryos co-cultured in 10% FCS(v/v)＋TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 72 hrs after bisection. In vitro survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The survival rate of bisected porcine embryos cultrued in 10% FCS＋TCM-199 media contaning PMSG, hCG, PMSG＋hCG, hCG＋$\beta$-estradiol 0 to 20 hrs and 20 to 40 hrs were 37.6% and 37.5%, 28.6% and 28.6%, 35.7% and 28.8%, 30.8% and 23.1%, 38.5% and 30.7%, respectively. The survival rate of bisected embryos co-cultured in TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells was significantly higher than that of non co-culture. 2. The survival rate of bisected porcine embryos co-cultured 10% FCS＋TCM-199 media containing oviductal epithelial cells 4 to 5 hrs and 20 to 24 hrs were 42.9% and 38.5%, respectively. 3. The survival rate of bisected porcine embryos co-cultured in 10% FCS＋TCM-199 media containing cumulus cells 4 to 5 hrs and 20 to 24 hrs were 40.0% and 35.7%, respectively.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 1997년도 제34차 추계 학술대회
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• pp.44-44
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• 1997

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2004년도 제47차 추계학술대회
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• pp.100-100
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• 2004

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2005년도 제49차 추계학술대회
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• pp.105.2-106
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• 2005

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.82-82
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• 2001

본 연구는 한우에 있어서 인공수정후 수태율증진을 위한 기초자료를 확보할 목적으로 성숙 미경산 한우의 발정동기화를 위해 CIDR(EAZI-BREED CIDR Plus, USA)를 7일간 질내에 삽입하였고, CIDR처리 제 6일째에 PGF2$\alpha$(lutlyase, USA)제제 5 $m\ell$을 근육주사하여 발정이 유기된 92%의 개체를 인공수정을 실시하였다. 수태율증진을 위해 무처리구를 두었고, hCG처리구(Chorulon, Intervet, Netherlands)는 2000IU를 근육 주사하였으며, CIDR처리구에는 Estradiol-17$\beta$의 캡슐을 제거한 CIDR를 수정후 7일부터 일주일간 질내에 삽입 처리하여 각 처리당 15두씩 총 45두의 한우를 실험에 공시하였다. 발정이 유기된 개체는 인공수정후 30일까지 2일 간격으로 채혈하여 혈중 Progesterone, IGF, IGF-II 및 cortisol 농도를 RIA법으로 분석하였으며 수정 후 60일 이후에는 직장검사법으로 임신감정을 실시하였다. 수정후 수태율은 대조구와 CIDR 처리구에서는 평균 62%였으나 hOG처리구는 73%로서 대조구에 비해 유의적으로 수태율이 높게 나타났다. 이때의 혈중 progesterone농도를 분석한 결과, 대조구에 CIDR 처리구의 수정후 7일째부터 증가하기 시작하여 제 12일째에서 평균 5.5ng/$m\ell$까지 증가하다가 그 이후 재발정 예정일일 21일째에 다소 감소되다가 다시 증가되어 일정한 수준으로 유지되었다 그러나 hCG처리구의 경우 수정후 5일부터 progesterone이 증가하다가 제 15일까지 약 10ng/$m\ell$까지 높은 progesterone이 유지되었다. 이와 같은 결과로 보아 hCG는 혈중 progesterone농도를 증가시킴으로써 난소내 임신황체의 progestorone분비 능을 촉진시키는 작용을 하는 것으로 사료되나, 한편 혈중 IGF I과 IGF-II농도는 대조구, hCG처리구 및 CIDR 처리구간의 차이가 없이 수정 후 제10일까지는 다소 높은 수준이었으나 그 이후 감소되는 현상으로 혈중 progesterone농도와는 부의 상관관계를 보였다. 따라서 IGF-I과 IGF-II는 혈중 progesterone의 농도를 인위적으로 조절하거나 황체의 progesterone분비기능을 직접 조절하는 역할은 하지 않는 것으로 사료되며, 특히 초기 임신의 수태율에 직접적으로 영향이 없는 것으로 사료된다. 또한 혈중 cortisol농도는 처리간의 차이는 확인할 수 없으나 임신과 비임신간의 혈중 cortisol농도는 유의적으로 차이가 있는 것으로 나타났다. 본 실험의 결과로 인공수정이나 수정란이식 및 체세포복제 수정란뿐만 아니라 형질전환 수정란의 이식후 수태율을 증진시켜 첨단기술의 조기정착을 위해 이식 후 7일째나 인공수정 후 7일째 hCG제제를 근육주사로 간단하게 처리함으로써 수태율을 크게 개선될 수 있으며 이것은 난소의 황체기능 즉, 황체로부터 충분한 progesterone을 분비할 수 있는 내분비적 환경을 제공하는 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제15권2호
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• pp.135-147
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• 1988

To compare the stimulation effect of the ratio in follicle stimulating hormone and luteinizing hormone in induction of multiple follicular growth, the serum $E_2$ level, the diameter of follicle, number of aspirated follicles and cleavage rate of in vitro fertilized preovulatory oocytes as well as the pregnancy rate were evaluated. Forty one patients with irreparable tubal disease were stimulated by hMG(n=24) or FSH/hMG(n=17) for the purpose of in vitro fertilization and embryo transfer. The following results were obtained. 1. Serum estradiol($E_2$) levels on the day of hCG administration were $921.0{\pm}353.3\;pg/ml$ in hMG group and $1272.9{\pm}1060.6\;pg/ml$ in FSH/hMG group. The serum $E_2$ value of hMG group was significantly lower than that of FSH/hMG group. 2. The diameter of leading follicle by ultrasonogram on the day of hCG administration were $16.2{\pm}2.0\;mm$ in hMG group and $16.2{\pm}2.6\;mm$ in FSH/hMG group. No significant difference of follicle diameter between two groups was demonstrated. 3. The number of follicles with diameter above 10 mm by sonogram on the day of hCG injection were $3.91{\pm}2.32$ in hMG group and $6.52{\pm}3.86$ in FSH/hMG group. There was significant difference of number of follicles between two groups, (p< 0.01). 4. The number of oocytes found per patient at aspiration were $2.59{\pm}1.00$ in hMG group and 3. $76{\pm}2.31$ in FSH/hMG group. There was significant difference of number of aspirated oocytes between two groups. (p< 0.05). 5. The detection rate of preovulatory oocyte at aspiration were 68.4%(39/57) in hMG group (n=22) and 77.6%(38/49) in FSH/hMG group (n=13). 6. The cleavage rate of preovulatory oocyte at 44 hours after insemination were 74.4%(29/39) in hMG group(n=22) and 81.6%(31/38) in FSH/hMG group (n=13). When only hMG was used, one pregnancy was established in 15 patients to whom 29 zygotes were transferred. And a full term normal female baby was delivered by elective cesarean section. In the FSH/hMG group, five pregnancies out of 9 transferred patients were confirmed by serum ${\beta}-hCG$. Two pregnancies were spontaneously aborted before the 6th week of pregnancy. One patient aborted her baby at the 18th week of pregnancy because of incompetent internal os of the cervix. Two patients delivered two full term babies by elective cesarean section. From the above findings, paralell with the increase in the ratio of exogenous follicle stimulating hormone to luteinizing hormone, an increase in oocyte recovery was observed as well as an improvements in pregnancy rate. It was concluded that FSH enrichment early in the follicular phase had a beneficial effect in the controlled ovarian hyperstimulation.