젖소 동결수정란의 비외과적 이식에 있어서 수정란의 상태 및 이시조건이 수태율에 미치는 영향

이은송,조충호,황우석,Lee, Eun-song,Jo, Choong-ho,Hwang, Woo-suk 대한수의학회 1989 大韓獸醫學會誌 Vol.29 No.3

This study was performed to investigate the effects of stage and quality of embryo, synchrony between donor and recipient and difficulty of transfer on pregnancy rate following non-surgical transfer of frozen-thawed bovine embryos. The results were as follows; 1. The overall pregnancy rate of this experiment was 63.4% and that of heifers(73.1%) was higher than that of cows(46.7%). 2. The pregnancy rates of recipients transferred with morulae, early blastocysts and blastocysts were 50.0%, 64.7% and 71. 4%, respectively. 3. The pregnancy rate of recipients transferred with good embryos(67.9%) was higher than that of recipients transferred with fair embryos(53.8%). 4. The pregnancy rates of embryos transferred to left and right uterine horn were 63.2% and 63.6%, respectively. 5. The pregnancy rate of recipients in estrous synchrony 0(76.2%) was higher than those of recipients in synchrony -1(55.6%) and +1(44.4%). 6. The pregnancy rate of recipients transferred with 2 embryos (71. 4%) was higher than that of recipients transferred with 1 embryo(61.8%). 7. The pregnancy rate of embryos transferred to uterine tip (72.0%) was higher than that of embryos transferred to uterine base(50.0%). 8. Ease of transfer was ranked to a scale of one to three on the basis of increasing difficulty. Transfers ranked as ease score 1 accounted for 77.8% of pregnancies and had higher pregnancy rate than ease score 2(66.7%) or 3(45.5%). 9. The pregnancy rate of recipients with excellent corpus luteum(CL) (70.0%) was higher than those of recipients with good CL(61.1%) or fair CL(61.5) %. In reviewing above results, it was considered that the factors such as embryo stage, embryo quality, estrous synchrony, corpus luteum quality, transfer site within uterus, recipient's parity and ease score affected the pregnancy rate after non-surgical transfer of frozen-thawed bovine embryos.

Effects of variation in the number and developmental stage of donor embryos and ovulation status of the surrogate mother on the efficiency of pig somatic cell cloning

Mi-Ryung Park,Jae Gyu Yoo,Chang-Gi Hur,Bo-Woong Sim,Myunghoo Kim,Jakyeom Seo,Byeong-Woo Kim,Byung-Wook Cho,Teak-Soon Shin,Seong-Keun Cho 한국동물생명공학회(구 한국동물번식학회) 2020 Journal of Animal Reproduction and Biotechnology Vol.35 No.3

This study investigated the effect of variation in the number of somaticcell- cloned embryos and their developmental stage at transfer on pregnancy, as well as the influence of the estrus status of recipient pigs on in vivo development of cloned porcine embryos after embryo transfer. For somatic cell nuclear transfer (SCNT), fibroblast cells were obtained from a male porcine fetus. Recipient oocytes were collected from prepubertal gilts at a local abattoir and then cultured. After SCNT, reconstructed embryos of different numbers and developmental stages were transferred into recipient pigs. The developmental stage of the cloned embryos and the number of transferred embryos per surrogate showed no significant differences in terms of the resulting cloning efficiency. However, the pregnancy rate improved gradually as the number of transferred cloned embryos was increased from 100- 150 or 151-200 to 201-300 per recipient. In pre-, peri-, and post-ovulation stages, pregnancy rates of 28.6%, 41.8%, and 67.6% and 16, 52, and 74 offspring were recorded, respectively. The number of cloned embryos and estrus status of the recipient pig at the time of transfer of the cloned embryo affect the efficiency of pig production; therefore, these variables should be particularly considered in order to increase the efficiency of somatic cell pig cloning.

체외수정란 이식시 수태율에 미치는 요인에 관한 연구

김성기,노상호,이은송,이병천,황우석,Kim, Sung-ki,Roh, Sang-ho,Lee, Eun-song,Lee, Byeong-chun,Hwang, Woo-suk 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.4

In the last few years, methods for in vitro culture of early embryo stages from oocytes matured and fertilized in vitro using suitable cell culture systems have been established. But the factors affecting pregnancy rates following transfer of bovine embryos produced in vitro were not evaluated enough. So this study was performed to investigate the effects of quality and stage of embryos, parity and Corpus Luteum quality of recipients on pregnancy rates following non-surgical transfer of bovine embryos produced in vitro. Oocytes aspirated from small antral follicles of ovaries obtained at a local slaughter house were matured, fertilized with frozen-thawed semen and co-cultured for 6-7 days by utilizing co-culture system with bovine oviduct epithelial cell in vitro. After co-culture, embryos were transfered to recipients on day 7 (estrus=day 0). Recipients were monitored by ultrasonic scanning method or observation for estrus and rectal palpation after 50 days from transfer. The results of this study are follows. 1. Of the 70 recipients, 70%(49 of 70) had not showed estrus sign between day 0 and day 50, but 22.9%(16 of 70) was diagnosed not pregnant. Therefore the overall pregnancy rate of this study was 47.1%(33 of 70). 2. The pregnancy rate of recipients transfered with excellent(66.7%) and good(54.5%) embryos were higher than that of recipients transfered with fair embryos(15.8%) (p<0.05). 3. The pregnancy rate of recipients transfered with morula, compacted morula, blastocyst and expanded blastocysts were 46.2, 55.0, 62.5 and 50.0%, respectively. 4. The pregnancy rates of recipients transfered to heifer and cow were 54.5 and 55.2%, respectively. 5. The pregnancy rates of recipients with CL score I, II(66.7, 63.6%) were higher than those of recipients with CL score III (10%), (p<0.05). Success of transfer of embryos produced in vitro depends on many variables. The important factors identified in this study were the quality of embryos and the CL score of recipient animals after non-surgical transfer of embryos matured, fertilized and cultured in vitro.

Technology Development of the Embryo Transfer and Production of Nuclear Transfer Embryo in Flower Deer and Elk

Jang-Hee Lee,Soon-Hwa Baek,Joo-Hyung Lee,Buom-Yong Ryu,Ki-Joong Kim,Yong-Hee Kim,Seong-Jae Park,Tai-Young Hur,Yeong-Hoon Jeong,Chang-Yong Choe,Ho-Sup Shim 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1

Artificial insemination and embryo transfer is one of the most important factors affecting to the production of fawn from deer nuclear transfer in the field of deer farms. This study* was conducted to establish the production technology of nuclear transfered embryo in deer. For estrus synchronization or superovulation tretments in flower deer and elk, each 10 does were inserted into the vagina for 14 days with CIDR (Pfizer New Zealand Ltd., NZ) for elk and Ring-CIDR (Bioculture Co., Ltd., Korea) for flower deer, and then those inserted devices were removed. The estrus synchronization of each 6 does were induced by the intramuscular injection of PGF2α (25 mg/head) and PG600 (hCG 200IU + PMSG 400IU, Intevet, Holland). Then, the superovulation of each 4 does of flower deer and elk was induced by additional injection of FSH (200 mg/ head) twice with an interval of 24 hours , respectively. Follicular oocytes were collected from each 2 does superovulated after 48 hours since the injection of PG600 and FSH. In the meantime, the ovarian response and the number of the collected ovarian follicles were investigated with the surgical operations. As a result, the average number of the collected ovarian follicles were 8.5 and 9.0 in flower deer and elk, respectively. The ovarian follicles collected from each two does were cultured in vitro for 48 hours with m-DMEM medium, and then the cell fusion was carried out after the nuclear transfer by the antler cell. As a result, 5 out of 18 ovarian follicles collected from 2 elk does were reached on the MII stage, but there was no generation resulting from the nuclear transferred embryos by the antler cell after enucleation. In 2 flower does, 7 out of 17 ovarian follicles were reached to the MII stage, but one of them was developed to parthenogenetic embryo as well despite a case of fusion from the nuclear transferred embryo. Embryos were collected in a surgical way on the 7th day after artificial insemination, numbers of average embryos collected were 2.5 and 3.0 in each 2 flower deer and elk does superovulated, respectively. The collected two embryos were transplanted to each 2 does synchronized. As a result, a head of fawn was produced from only one elk doe, where as a head of fawn were delivered from one out of 4 elk does artificial inseminated. Given these findings, we consider that more or less of problems might have occurred in vitro culture system of ovarian follicles in the production of nuclear transfered deer embryos. In addition, the greatest reason why both the aetificial insemination and embryo transfer failed was considered attributable to stress due to anesthesia and catching.

체외수정시술시 배아이식 후 배아이식도관 말단부에서의 미세균주 배양율과 임상적 임신율과의 관계

이경진,배상욱,김정연,김진영,이병석,박기현,조동제,송찬호,Lee, Kyoung-Jin,Bai, Sang-Wook,Kim, Jeong-Yeon,Kim, Jin-Young,Lee, Byung-Seok,Park, Ki-Hyun,Cho, Dong-Jae,Song, Chan-Ho 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.3

Objective: To evaluate incidence of microbial growth from the tip of the embryo transfer catheter after embryo transfer in relation to clinical pregnancy rate following in-vitro fertilization and embryo transfer. Method: This study was performed prospectively at the time of transcervical embryo transfer following conventional in-vitro fertilization and intracytoplasmic sperm injection procedures. Sixty three patients were enrolled in this study. Microbiological cultures were performed on endocervical swabs and embryo transfer catheter tips. Results: Positive microbial growths were observed from endocervical swabs in 45 (71.4%) women and from catheter tips in 30 (47.6%) women. There was no statistically significant difference seen in the mean number of oocytes fertilized or number and grade of embryos transferred between the group of patients without growth and the group of patients with positive microbial growth from catheter tips. The clinical pregnancy rate were 30.3% in the group of patients without growth and 13.3% in the group with positive microbial growth from catheter tips. This difference in clinical pregnancy rates was statistically significant. Conclusion: Our finding is that microbial contamination at embryo transfer may influence implantation rates. The major questions arising from our finding are whether eradication of endocervical micro-organisms is possible and whether their eradication will improve implantation rates.

Laparoscopy vs. laparotomy for embryo transfer to produce transgenic goats (Capra hircus)

Sang Tae Shin,Sung Keun Jang,Hong Suk Yang,Ok Keun Lee,Yhong Hee Shim,Won Il Choi,Doo Soo Lee,Gwan Sun Lee,Jong Ki Cho,Young Won Lee 대한수의학회 2008 JOURNAL OF VETERINARY SCIENCE Vol.9 No.1

This study was performed to produce transgenic Korean native goat (Capra hircus) by laparoscopic embryo transfer (ET) to overcome the limitations of ET performed by laparotomy. Transgenic embryos were produced by DNA pronuclear microinjection of in vivo zygotes. The recipient goats were synchronized for estrus by using an introvaginal progesterone devices as a controlled internal drugreleasing insert (CIDR) for 13 days and injection of 400 IU PMSG 48 h before removal of the insert. Embryos were transferred on day 3 and 4 after removal of the insert. Recipient goats were deprived of feed for 48 h, then suspended in a laparotomy cradle at an angle of 45o. After obtaining a sufficient pneumoperitoneum, the laparoscope and forceps were inserted abdominally through 5 mm trocar sleeves. Examination of the ovaries and uterus was performed and then 213 embryos were transferred into the oviducts via the infundibula of 76 recipient goats. To compare pregnancy rates, ET was also performed by laparotomy in 82 recipient goats. The pregnancies in the recipient goats were diagnosed by ultrasound on day 30 after embryo transfer. The pregnancy rate with laparoscopic ET was significantly higher than with ET performed by laparotomy (46.1% vs. 28.6%, p < 0.05). In addition, the pregnancy rates were compared between ovulated and non-ovulated ovaries of the recipient goats in the laparoscopic ET group. No significant difference was observed between the pregnancy rates of ovulated and non-ovulated ovaries (41.3% vs. 33.3%, p < 0.05) suggesting that ET may also be possible in non-ovulated recipients through artificial rupture of Graafian follicles. These results suggest that laparoscopic ET is a highly efficient method for the transfer of goat embryos. This study was performed to produce transgenic Korean native goat (Capra hircus) by laparoscopic embryo transfer (ET) to overcome the limitations of ET performed by laparotomy. Transgenic embryos were produced by DNA pronuclear microinjection of in vivo zygotes. The recipient goats were synchronized for estrus by using an introvaginal progesterone devices as a controlled internal drugreleasing insert (CIDR) for 13 days and injection of 400 IU PMSG 48 h before removal of the insert. Embryos were transferred on day 3 and 4 after removal of the insert. Recipient goats were deprived of feed for 48 h, then suspended in a laparotomy cradle at an angle of 45o. After obtaining a sufficient pneumoperitoneum, the laparoscope and forceps were inserted abdominally through 5 mm trocar sleeves. Examination of the ovaries and uterus was performed and then 213 embryos were transferred into the oviducts via the infundibula of 76 recipient goats. To compare pregnancy rates, ET was also performed by laparotomy in 82 recipient goats. The pregnancies in the recipient goats were diagnosed by ultrasound on day 30 after embryo transfer. The pregnancy rate with laparoscopic ET was significantly higher than with ET performed by laparotomy (46.1% vs. 28.6%, p < 0.05). In addition, the pregnancy rates were compared between ovulated and non-ovulated ovaries of the recipient goats in the laparoscopic ET group. No significant difference was observed between the pregnancy rates of ovulated and non-ovulated ovaries (41.3% vs. 33.3%, p < 0.05) suggesting that ET may also be possible in non-ovulated recipients through artificial rupture of Graafian follicles. These results suggest that laparoscopic ET is a highly efficient method for the transfer of goat embryos.

Deposition of Mucin Coat on Rabbit Embryos Cultured In Vitro Following Oviductal Transfer

Joung S. Y,Yang J. H,Im K. S.,Lee S. H.,Park C. S.,Jin D. I. 한국동물생명공학회(구 한국동물번식학회) 2004 Reproductive & developmental biology Vol.28 No.3

Mucin coat is deposited on the embryos during passage through the oviduct in rabbit. When in vitro cultured blastocysts were transferred to the recipients, the lack of mucin coat might account in part for failure of pregnancy after transfer. The present study were carried out to investigate whether deposition of mucin coat were induced when in vitro cultured blastocysts were transferred to recipients. At 19 ~20 hours post-coitus one-cell embryos were collected by flushing oviducts. These embryos cultured for 72 hours were reached to blastocyst stage. And these blastocysts were transferred to the oviduct of asynchronized (one day later than the donors) and synchronized recipient. To confirm deposition of the mucin coat, blastocysts transferred to the oviduct were recovered at 24 and 48 hours after the transfer. Fifty eight percent of blastocysts recovered from uterus of asynchronous recipient at 24 hours after transfer and 92.9% of blastocysts recovered from uterus of synchronous recipient were 0~10 ㎛ of mucin coat thickness. And 11.8% of blastocysts of asynchronized recipients and 7.1% of blastocysts from asynchronized recipients were in 11~20 ㎛ of mucin coat thickness. When blastocysts were recovered from uterus at 48 hours after transfer, 87.0% of blastocysts from asynchronized recipients and 5.9% of blastocyst from synchronized recipients were in 0~10 ㎛ of mucin coat thickness. And 76.5% of blastocysts of synchronized recipients and 4.4% of blastocysts from asynchronized recipients were in 11~20 ㎛ of mucin coat thickness. From these results it is speculated that the low implantation rate of in vitro cultured rabbit blastocysts transferred to oviduct of recipient was caused by high degeneration of the embryo after transfer and inappropriate deposition of mucin coat.

Viability of Somatic Cell Nuclear Transfer Embryos following Embryo Transfer in Korean Native Striped Cattle(Bos Namadicus Falconer, Chikso)

Dae-jin Kwon,Joo-hee Park,Hwan-Sub Hwang,Yeon-Soo Park,Choon-Keun Park,Boo-Keun Yang,Hee-Tae Cheong 한국동물생명공학회(구 한국동물번식학회) 2007 Reproductive & developmental biology Vol.31 No.4

This study was conducted to examine the viability of Korean native striped cattle (Bos namadicus Falconer, Chikso) clone embryos after embryo transfer. Chikso somatic cell nuclear transfer (SCNT) embryos were produced by fusion of ear skin cells derived from a female Chikso with enucleated oocytes matured in vitro for 18-24 hr. After in vitro culture of SCNT embryos for 7 to 8 days, fresh or vitrified blastocysts derived from SCNT were transferred into a uterine horn of recipient cows. Fifteen of total 43 recipients were pregnant at Day 50 and 4 recipients were maintained to term. Three IVF-derived calves and 1 clone Chikso calf were born. Pregnancy rate was higher when fresh embryos were transferred to recipients compared to vitrified embryos, but development to term was not different between both groups. The clone Chikso calf died at 5 days after birth due to the fullness of amniotic fluid in rumen and the infection of umbilical cord. The result of the present study shows that clone Chikso calf can produced from the embryo transfer of SCNT embryos, however, solution of abortion problem is necessary to improve the cloning efficiency.

Improved pregnancy rate and sex ratio in fresh/frozen in vivo derived embryo transfer of Hanwoo (Bos taurus coreanae) cows

Jihyun Park,Wonyou Lee,Islam M. Saadeldin,Seonggyu Bang,Sang Hoon Lee,이준구,Jong Ki Cho 한국축산학회 2023 한국축산학회지 Vol.65 No.4

This study aimed to assess the effects of embryonic developmental stage, quality grade, and fresh or frozen/thawed conditions on the pregnancy rate and sex ratio of live offspring in Hanwoo (Bos taurus coreanae) cows. The quality and developmental stage of in vivo-derived (IVD) transferred embryos were evaluated using the standard criteria of the International Embryo Technology Society. The recipient cows were synchronized using conventional (estradiol benzoate and progesterone) protocols before embryo transfer. Embryos were transferred to 297 cows, and pregnancy was monitored for 60–70 days after embryo transfer. The pregnancy rates of fresh and frozen/thawed embryos were 56.90% and 52.49%, respectively. Pregnancy rates varied according to embryo quality (56.18% for grade 1 vs. 36.67% for grade 2). Pregnancy rates also varied by developmental stage and cryopreservation (67.86% vs. 63.49% for stage 4-1, 64.00% vs. 54.72% for 5-1, and 50.00% vs. 47.83% for 6-1, in fresh embryos vs. frozen/thawed embryos, respectively). For stage 7-1, the pregnancy rates were 72.73% for fresh embryos and 20.00% for frozen/thawed embryos. In 66 fresh embryos, the sex ratio of live offspring was 5:5, whereas it was 4(female):6(male) for frozen/thawed embryos among the 95 frozen/thawed embryos. The miscarriage rate was approximately 3% higher for frozen/thawed embryos than for fresh embryos (18.1% for fresh vs. 21.1% for frozen). Seasonal fertility rates were 33.3% in spring, 55.67% in summer, 52.8% in autumn, 60.0% in winter. The following male-to-female ratios were observed in different seasons: 6.7:3.3 in spring, 4.0:6.0 in summer, 5.5:4.5 in autumn, and 3.3:6.7 in winter. The current data revealed no significant differences in pregnancy rates between fresh and frozen/thawed IVD embryos. However, there was a lower pregnancy rate with advanced-stage frozen/thawed embryos (stage 7-1). The current study provides comprehensive results for the better optimization of embryo transfer in Hanwoo cattle to obtain the desired fertility rate, pregnancy rate, and sex ratio of calves. These results provide important insights into the factors that influence the viability and success of IVD embryo transfer in Hanwoo cows and may have practical applications for improving breeding programs and reducing production costs.

Viability of Somatic Cell Nuclear Transfer Embryos following Embryo Transfer in Korean Native Striped Cattle (Bos namadicus Falconer, Chikso)

Kwon, Dae-Jin,Park, Joo-Hee,Hwang, Hwan-Sub,Park, Yeon-Soo,Park, Choon-Keun,Yang, Boo-Keun,Cheong, Hee-Tae The Korean Society of Animal Reproduction 2007 Reproductive & developmental biology Vol.31 No.4

This study was conducted to examine the viability of Korean native striped cattle (Bos namadicus Falconer, Chikso) clone embryos after embryo transfer. Chikso somatic cell nuclear transfer (SCNT) embryos were produced by fusion of ear skin cells derived from a female Chikso with enucleated oocytes matured in vitro for 18-24 hr. After in vitro culture of SCNT embryos for 7 to 8 days, fresh or vitrified blastocysts derived from SCNT were transferred into a uterine horn of recipient cows. Fifteen of total 43 recipients were pregnant at Day 50 and 4 recipients were maintained to term. Three IVF-derived calves and 1 clone Chikso calf were born. Pregnancy rate was higher when fresh embryos were transferred to recipients compared to vitrified embryos, but development to term was not different between both groups. The clone Chikso calf died at 5 days after birth due to the fullness of amniotic fluid in rumen and the infection of umbilical cord. The result of the present study shows that clone Chikso calf can produced from the embryo transfer of SCNT embryos, however, solution of abortion problem is necessary to improve the cloning efficiency.