• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2000년도 수정란의 위생적 처리·검사 및 특수가축의 수정란이식 기술 심포지움
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• pp.64-75
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• 2000

During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period(1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor(hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

• 한국수정란이식학회지
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• 제12권3호
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• pp.301-306
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• 1997

The present study was carried out to investigate effect of FSH -P dose and energy level on normal embryo production after superovulation in Hanwoo. The results obtained were as follows ; 1. There was a significant effect of dose of FSH-P on normal embryo production in Hanwoo(P$\pm$5.9), 40(4.9i5.7), 50mg(2.2$\pm$2.6). 2. The plasma P$_4$ levels on the first treatment day were higher group( >4ng /ml) than lower group( <=4ng /ml), produced significicantly(P<0.05) higher number of normal embryos. 3. There was a significant effect of energy level on normal embryo production in Hanwoo(P$\pm$6.0), number of normal embryos were higher than TDN 70%(5.1$\pm$6.5) and TDN 130%(4.4$\pm$2.6) 4. The donor returned to normal estrus after superovulation were 44.8, 28.4 and 29.9 days by TDN 70, 100 and 130%, respectively.

• 한국수정란이식학회지
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• 제17권1호
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• pp.23-32
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• 2002

본 연구는 기존의 수정란 이식기술을 다각적으로 분석하고 개선하여 한우 체내수정란의 생산 및 이식기술 체계를 확립하기 위하여 수행하였으며, 결과를 요약하면 다음과 같다. 1. 배란에 미치는 FSH analogue 간의 유의성은 인정되지 않았으나 수정률, 이식 가능 수정란 및 동결수정란 생산에는 처리간에 유의성(P<0.05)이 인정되었다. 특히, Super-OV는 Foll-tropin-V 및 Embryo-S에 비해서 이식 가능 수정란의 생산율이 유의적(P<0.01)으로 낮았다. 2. 호르몬의 투여시기에 따라 공란우의 배란정도 (P<0.1), 수정률, 이식 가능 및 동결 가능 수정란의 생산(P<0.05)은 각각 유의적인 차이를 나타냈다. 특히, 발정주기 11일차의 호르몬투여는 적합하지 않은 결과를 보였다. 3. 다배란처리후 황체의 등급에 따른 수정란의 수는 좌측 또는 우측에서 6개 이상의 황체를 보인 A등급(8.06∼9.49개)의 경우가 B, C 및 D등급(2.93 ∼6.28개)보다 유의적(P<0.01)으로 높았다

• 한국수정란이식학회지
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• 제12권1호
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• pp.57-66
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• 1997

This study was carried out to establish an effective and practical system for commercialization of embryo production techniques by analyzing several factors influencing in vivo embryo production on days and seasons of flushing in Korean native cattle. In vivo embryos were flushed 226 times from 128 donors. The results obtained for the factors influencing in vivo embryo production on days and seasons of flushing were as follows :1.The percentages of fertilized, transferable and freezable embryos by seasons were significantly different in both FSR-P and SUPER-OV(P<0.01). The percentages of them were highest in sunrrner with FS H-P and highest in autumn with SUPER -OV.2. The production of transferable and freezable embryos by flushing days was highest in 8 days with FSH-P, and there was no difference between 7 and 8 days for SUP ER-OV. 3. The failure rates of recovery were 17.0% in SUPER-OV and 21.2% in FSH-P, respectively. The donors superovulated but failed recovery were 8.5% in SUPER-OV and 12.9% in FSH-P, respectively. Nonsuperovulated donors was 8.4% and donors giving less than 2 eggs at recovery was 8.4% in both FSH -P and SUPER-OV 4. The donors returned to normal estrus after superovulation were 34.1% after 1 cycle,39.4% after 2 cycles, and 16.7% after 3 cycles by FSH-P, respectively. For SUPER-OV, they were 55.3, 33.0 and 9.6%, respectively. Generally, normal estrus after the treatment of superovulation was earlier and the occurrence of ovarian cyst was also lower in SUP ER-OV than in FSH-P.5.The percentages of blastocyst in embryos flushed at 7~8 days after estrus were 21. 9% and 54.3% in FSH -P and SUPER-OV, respectively. The development of embryos was faster in SUPER-OV than in FSH-P.(Key words : in vivo embryo, flushing days, superovulation, FSH-P, SUPER-OV)

• Asian-Australasian Journal of Animal Sciences
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• 제19권2호
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• pp.171-175
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• 2006

The aim of this in vitro study was to test the effect of microinjection (Mi) of foreign gene into the rabbit egg pronucleus and epidermal growth factor (EGF) addition on the blastocyst rate, the cell number and the diameter of embryos, and to determine possible relationships between embryo cell number and embryo diameter. Blastocyst rate was significantly decreased in gene- Mi (G-Mi/E0) group (63.1%) comparing to intact ones (83.5%, $p_1$<0.05). The addition of EGF at 20ng/ml (G-Mi/E20) or 200 ng/ml (GMi/ E200) to gene-Mi embryos did not affect blastocyst rate (65.6 and 55.2% resp.). As a control for Mi, the eggs were microinjected with the same volume of phosphate-buffered solution (PBS-Mi) instead of the gene construct solution. Cell numbers and embryo diameters were measured from embryo images obtained on confocal laser scanning microscope. Bonferroni-modified LSD test showed that the embryo cell number in PBS-Mi group was significantly lower ($p_1$<0.05) and in gene-Mi group was tended to decrease compared with intact embryos. Embryo diameter was not different among experimental groups. No effect of EGF given at any doses both on the cell number and embryo diameter was found. A positive correlation between cell number and embryo diameter was observed in all groups of embryos. Since embryo diameter was not changed under the influence of Mi or EGF addition in this study, this seems to be more conservative characteristics of the embryo morphology. These results suggest that the pronuclear microinjection compromises developmental potential of embryos, decreasing blastocyst rate and embryo cell number, whilst embryo diameter is not affected. No effects of EGF on studied parameters were confirmed. Declined quality of Mi-derived embryos is caused by the microinjection procedure itself, rather than by the gene construct used.

• 한국수정란이식학회지
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• 제33권4호
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• pp.265-270
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• 2018

Up-to-date artificial insemination (AI) using frozen sperm consider as the most widely using technology for improvement of Korean Native Cow (Hanwoo) embryo production. However, it is time consuming, required at least 15~20 years to make more than 6 generations, and their offspring number is limited. To overcome such limitations, superovulation and in vitro fertilization have been developed. For superovulation, the number of produced embryos are not enough for commercialization and donor cows need rest period. This led to use of slaughterhouse ovary for in vitro fertilization, but it is impossible to repeat the collection from the same individual and it only can improve the genetic merits of offspring for one generation. Production of embryos using Ovum Pick-Up (OPU) technique, where oocytes can be repeatedly collected from living elite donor, might overcome these limitations. In this study, we investigated the possibility of using OPU technique from donors at different age and different session periods for mass-embryo-production. Oocytes were collected from 26 donor cows twice per week, 3 - 4 months per year, between 2013 and 2016. Results showed that, the average number of embryo produced in first year used donor was significantly higher than that in second year used donor ($3.89{\pm}2.85$ vs $3.29{\pm}2.70$), however, there was no significant difference between third year used donor ($3.51{\pm}3.32$) and other groups. Taken together, our data showed that repeated using of donor up to three years is possible for in vitro embryo mass-production. Moreover, OPU can be used as suitable embryo producing technique for livestock breed improvement.

• 한국동물생명공학회지
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• 제37권2호
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• pp.80-86
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• 2022

The ability to determine the sex of bovine embryos before the transfer is advantageous in livestock management, especially in dairy production, where female calves are preferred in milk industry. The milk production of female and male cattle benefits both the dairy and beef industries. Pre-implantation sexing of embryos also helps with embryo transfer success. There are two approaches for sexing bovine embryos in farm animals: invasive and non-invasive. A non-invasive method of embryo sexing retains the embryo's autonomy and, as a result, is less likely to impair the embryo's ability to move and implant successfully. There are lists of non-invasive embryo sexing such as; Detection of H-Y antigens, X-linked enzymes, and sexing based on embryo cleavage and development. Since it protects the embryo's autonomy, the non-invasive procedure is considered to be the safest. Invasive methods affect an embryo's integrity and are likely to damage the embryo's chances of successful transformation. There are different types of invasive methods such as polymerase chain reaction, detection of male chromatin Y chromosome-specific DNA probes, Loop-mediated isothermal amplification (LAMP), cytological karyotyping, and immunofluorescence (FISH). The PCR approach is highly sensitive, precise, and effective as compared to invasive methods of farm animal embryonic sexing. Invasive procedures, such as cytological karyotyping, have high accuracy but are impractical in the field due to embryonic effectiveness concerns. This technology can be applicable especially in the dairy and beef industry by producing female and male animals respectively. Enhancing selection accuracy and decreasing the multiple ovulation embryo transfer costs.

• 한국수정란이식학회지
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• 제14권1호
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• pp.39-46
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• 1999

To establish the optimal culture systems for production of transferable embryos in Korean Cattle, pregnancy rates of IVF-derived blastocysts according to different culture media, culture method and culture duration were compared. Development of IVF-derived embryos to blastocysts was most effective in YS medium group co-cultre with cumulus cells. Blastocysts cultured for 6 to 8 d in vitro showed higher hatching rate and good quality. Pregnancy rates after transfer of IVF-derived blastocysts cultured for 7 or 8 d were high. Through our experiments, it is considered that improvement of culture media and culture method is necessary for mass production of blastocysts with excellent of good quality in Korean Cattle.

• 한국수정란이식학회지
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• 제8권1호
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• pp.37-44
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• 1993

Nuclear transplantation techinque has been found to be the most potential and efficient method for producing a large number of genetically identical animals from a single embryo. The technical development of nuclear transplantation in mammals and its application to the production of cloned animals were reviewed. For the efficient and successful production of cloned embryos by nuclear transplantation, selection and micromanipulation of recipient eggs or embryos as capacious recipient cytoplasm, and benefitial preparation of multiple totipotent embryonic cells as donor nuclei, and also fusion technique are very critical. Recent works approaching to these critical points were introduced and discussed.

• 한국수정란이식학회지
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• 제27권2호
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• pp.71-80
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• 2012

Biotechnologies for cloning animals and in vitro embryo production have the potential to produce biomedical models for various researches. Especially, pigs are a suitable model for xenotransplantation, transgenic production and various areas of reproductive research due to its physiological similarities to human. However, utilization of in vitro-produced embryos for transfer remains limited. Despite improvement over past few decades, obstacles associated with the production of good quality embryos in vitro still exist which limit the efficiency of cloning. One of major problems includes improper in vitro maturation (IVM) and culture (IVC). Oxidative stress caused from in vitro culture conditions contributes to inadequate IVM and IVC which leads to poor developmental competence of oocytes, failure of fertilization and embryo development. To reduce the oxidative stress, various antioxidants have been used to IVM and IVC system. However, limited information is available on the effects of resveratrol on livestock reproductions. Resveratrol is a polyphenolic natural product and well known as an antioxidant in foods and beverages (e.g. in grapes and red wine). Resveratrol is known to be cardioprotective, anticarcinogenic, anti-inflammatory, antioxidant and antiapoptotic. This paper will review the effects of resveratrol on in vitro maturation of oocytes and embryo development.