• Asian-Australasian Journal of Animal Sciences
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• 제17권4호
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• pp.453-459
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• 2004

Chick embryos from stage 14 to stage 31 were studied by means of serial section and light microscopy in order to learn the relationship between the settlement sites of the primordial germ cells (PGCs) and the forming genital ridge. The results showed that: when embryo hatched for 53-56 h, the PGCs reached the coelomic epithelial tissue where gonad would be formed, meanwhile the epithelial tissue began thicker before the PGCs reached. Before stage 19, the final region the PGCs arrived was the thickened portion of the coelomic epithelium, the glycogen in the PGCs cytoplasm maintenance remained unchanged. However at the 3.5-5th hatching day, the glycogen in the PGCs cytoplasm reduced gradually. On the 6th hatching day, the gonad of the embryo appeared the feature of ovary, and the glycogen in the PGCs cytoplasm reduced further. On the 7th hatching day, the differentiation of ovary or testis was obvious and the glycogen in the PGCs cytoplasm later disappeared.

• 한국수정란이식학회지
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• 제27권3호
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• pp.171-174
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• 2012

Various small molecules can be used to control major signaling pathways to enhance stemness and inhibit differentiation in murine embryonic stem cell (mESC) culture. Small molecules inhibiting the fibroblast growth factor (FGF)/ERK pathway can preserve pluripotent cells from stimulation of differentiation. In this study, we aimed to evaluate the effect of pluripotin (SC-1), an inhibitor of the FGF/ERK pathway, on the colony formation of outgrowing presumptive mESCs. After plating the zona pellucida-free blastocyst on the feeder layer, attached cell clumps was cultured with SC-1 until the endpoint of the experiment at passage 10. In this experiment, when the number of colonies was counted at passage 3, SC-1-treated group showed 3.4 fold more mESC colonies when compared with control group. However, after passage 4, there was no stimulating effect of SC-1 on the colony formation. In conclusion, SC-1 treatment can be used to promote mESC generation by increasing the number of early mESC colonies.

• 한국수정란이식학회지
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• 제32권3호
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• pp.81-86
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• 2017

The Somatic cell nuclear transfer (SCNT) method can be applied to various fields such as species conservation, regenerative medicine, farming industries and drug production. However, the efficiency using SCNT is very low for many reasons. One of the troubles of SCNT is that it is highly dependent on the researcher's competence. For that reason, four somatic cell nuclear injection methods were compared to evaluate the effect of hole-sealing process and existence of cytochalasin B (CB) on efficiency of murine SCNT protocol. As a results, the microinjection with the hole-sealing process, the oocyte plasma membrane is inhaled with injection pipette, in HCZB with CB was presented to be the most efficient for the reconstructed in SCNT process. In addition, we demonstrated that the oocytes manipulated in Hepes-CZB medium (HCZB) with CB does not affect the developmental rate and the morphology of the blastocyst during the pre-implantation stage. For this reason, we suggest the microinjection involving hole-sealing in HCZB with CB could improve SCNT process efficiency.

• Journal of Plant Biotechnology
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• 제42권4호
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• pp.376-379
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• 2015

본 연구는 낙엽송의 배발생조직 증식 및 체세포배 유도에 영향하는 유기질소원(${\small{L}}$-Glutamine 및 casein hydrolysate, CH), 식물생장조절물질(옥신 및 사이토키니류)의 종류 및 농도의 효과를 평가하기 위해 수행되었다. 배발생조직 증식 비교에서 1,000 mg/L ${\small{L}}$-Glutamine 첨가 시 2주 혹은 4주 모두 가장 높은 조직생중량을 보였고 그 외 처리구에서는 대체로 유사하였다. 체세포배 유도에서는 0.2 mg/L IBA 첨가 시 가장 높은 426.3개를 유도하여 가장 효과적이었으나 0.2 mg/L BA 혹은 Kinetin처리구에서는 전혀 체세포배가 유도되지 않았다. IBA 농도 별 비교에서는 0.2 mg/L(303개)농도가 가장 좋았으며, 1.0 mg농도에서도 281개를 유도하여 체세포배 유도에 효과적이었으나 5.0 mg/L 첨가 시 109.3개로 가장 저조하였다.

• 한국수정란이식학회지
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• 제24권4호
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• pp.289-292
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• 2009

Mongolia has 80% livestock of total agriculture industry, 170,000 farms are engaged, 2,500,000 of cows that were beef and dairy cows are raised. Despite of Mongolian has great application with milk, there are not clear differences between cow and dairy cattle, and the production of milk is also low. But the milk suppliers are varied (horse, sheep, goat, etc), so that the total milk production is 500 thousand ton per year. It's really considerable to improve the breed of owing to many problems with big differences among milk qualities. For carrying out for first year project, artificial insemination project was operated with 3rd grade Holstein semen that were imported from S. Korea, and initiation and field training were also carried out through appropriate AI technique we developed for Mongolia environment. Local information research and MOU conclusion were done with professor D. Altangerel in May $10^{th}{\sim}13^{th}$, 2009, and development for AI technique and AI equipments were supplied for Mongolia breeding and natural environment in July $10^{th}{\sim}17^{th}$ in 2009. All cows were treated by synchronization for AI. To do this, $PGF_{2\alpha}$ injection were treated for luteal phase cow, if it wouldn't work, try again after 11 days. After confirmation of estrus, AI and AI training were carried out with sperm injection in the uterus or cervix by rectum-vagina method which is common worldwide, the most effective artificial insemination technique. If cows were return to next estrus cycle, second AI was carried out about approximately 21 days after artificial insemination. After 2 months, all cows not showing return estrus should be taken pregnancy test. Every pregnant cow will be cared thoroughly. Total 48 cows administrated by $PGF_{2\alpha}$ for synchronization and after 48 hours 45 cows (93.8%) showing estrus were detected and then artificial inseminate them within who 8 cows (27.8%) showed return estrus. Therefore, Using $PGF_2{\alpha}$ for synchronization is effective to use for Mongolia breeding conditions. There are possibility of base for food production after all, including increase of livestock production in Mongolia by improvement of breeding cow with AI and embryo transfer project.

• 한국수정란이식학회지
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• 제23권3호
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• pp.217-222
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• 2008

In previous studies, we reported that sow which was transferred OPS-freezing embryos not able to deliver a piglet (Kim et al, 2004). This study was conducted to investigate a possibility of gilt as recipients which produce piglets after transfer of OPS-freezing embryos. All transferred embryos were prepared by in vitro production (IVP) system. In vitro culture (IVC) medium used glucose-free NCSU23 supplemented with 5mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$. From day 3 of IVC, 10% fetal bovine serum albumin was added to the culture medium. In preparing of freezing embryos, embryos were treated with 7.5 $\mu g/ml$ cytochalasin-B for 30 min and centrifuged at $13,000{\times}g$ for 13 min. And then, embryos were exposed sequentially to an ethylene glycol (EG) solution, aspirated into open pulled straw (OPS), and plunged or thawed into the liquid nitrogen. In embryo transfer (ET), we used two kinds of type (surgical method vs. non-surgical method). In surgical method of embryo transfer, $55\sim65$ embryo were transferred in both uterine horn of two recipient gilts by plastic straw. Non-surgical method which is like artificial insemination was performed on three gilts. Each 140 frozen embryos were transferred to two gilts and 40 fresh embryos to one gilt. Pregnancy establishment was shown one recipient at 45 days after ET. However, the one recipient was also aborted at 58 days after ET. These results suggest that gilts can be considered as a candidate of recipients for OPS-freezing embryo transfer.

• Asian-Australasian Journal of Animal Sciences
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• 제11권4호
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• pp.398-403
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• 1998

The objective of the present study was to examine the developmental rates, and the stage of development in relation to time since fertilization, of in vitro produced buffalo embryos. Buffalo cumulus-oocyte complexes obtained from slaughterhouse ovaries were matured and fertilized in vitro. The fertilized oocytes (n = 248) were then co-cultured with buffalo oviductal epithelial cells and evaluated for the developmental stages on Days 2, 4, 6, 7, 8, 9 and 10 post-insemination. The peak of 4-cell stage embryos was observed on Day 2 (63.7 %), whereas Day 4 was marked by peaks of 6-8-cell stage embryos (20.9%) and 16-cell stage embryos to early morulae (50%). On Days 6, 7, 8, 9, and 10 post-insemination, 49.5, 48.3, 38.3, 33.8 and 33.4% embryos were found to be at morula/compact morula stages, 8.8, 12.5, 25.4, 6.0 and 1.2% at early blastocyst/blastocyst stages, 0, 6.8, 7.2, 15.3 and 2.0% at expanded blastocyst stage and 0, 1.6, 4.8, 19.3 and 38.5% hatching/hatched blastocyst stages, respectively. The peaks of early blastocyst/blastocyst, expanded blastocyst and hatching/hatched blastocyst stages were observed on Days 8, 9 and 10, respectively. The percentages of oocytes which initially became arrested and subsequently degenerated were 3.6, 4.8, 10.4, 14.5, 21.3 and 24.5% on Days 4, 6, 7, 8, 9 and 10 post-insemination, respectively.

• 한국작물학회지
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• 제51권spc1호
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• pp.237-241
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• 2006

국내육성 유채품종 간 반수체식물 생산성을 비교하였다. 화아로부터 분리된 소포자를 13%의 sucrose, $0.05mg/{\ell}$ BA 와 $0.5mg/{\ell}$ 의 NAA가 첨가된 NLN 배지에서 배양하였다. 반수체 배의 생산에는 유전형이 중요한 요인이였으며, 탐라유채가 가장 높은 반수체 배 생산능을 보여 화아당 176개의 반수체 배가 발생하였으나, 한라유채와 영산유채는 배 생산은 물론 세포분열도 관찰되지 않았다. 발생한 반수체 배를 NLN배지에서 현탁배양하여 Multilobe abnormal embryos를 형성시켰으며, 계속하여 생장조절제가 첨가되지 않은 MS고체 배지에 치상 배양하여 반수체식물체를 유도 하였다. 재생된 반수체 식물체는 성공적으로 순화되었다.

• 한국수정란이식학회지
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• 제1권1호
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• pp.35-49
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• 1986

Recipients are an integral part of embryo transfer and they are expensive to maintain as a good recipient. Recipient management is one of the most important components in a successful embryo transfer program. Management includes selection and subsequent care of the animals. A good recipient is basically on "open" cows or heffers whose reproductive tract is capable of receiving one or two embryos and incubating it to term. Potential recipients should be always be healthy and cycling normally ranging from 18 to 23 days. A thorough veterinary examination is recommended for candidate of recipients and cattle for questionable health should be eliminated from the recipient herd. Age and size of recipients are particularly important considerations when heifers are used, because of most embryos available for transfer are from large dams and sires. Body condition can influence a recipient's production, reproduction and health. Obese and underconditioned cattle should be avoided for use. Transfer of fresh embryos especially requires precise synchronization of donors and recipients. For estrus synchronization, PGF$_2$$\alpha$ is injected twice 10 to 12 days apart and short4erm progestagen treatment is applied to potential recipient cattle by coil into vagina (PRID) or ear implant (Synchro-Mate-B). The highest pregnancy results are achieved in recipients at exact synchrony with donors or 12 to 24 hr earlier than donors. Estrus detection is a major factor in breeding efficiency. High accuracy can be achieved by use of heat mount detection alds or by obserbing cattle for 30-minute peroids 3 times daily. Assay progesterone in milk can be used to discrIminate between pregnant and nonprenant recipients. Rectal palpation on day 35 to 70 after is an accurate and safe method of pregnancy diagnosis. Embryonic mortality in recipients may be associated with factors such as high environmental temperature and nutritional or lactational stress in early lactation period. Achievement of short calving interval requires concentrated management activity during the first 90 days following calving. Acceptable candidate for a recipient should be routinely vaccinated for infectious diseases. Proper nutritional programs according to NRC requirements and body condition scoring system for recipient cattles are vital to the ultimate success of an embryo transfer program.r program.

• Asian-Australasian Journal of Animal Sciences
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• 제28권11호
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• pp.1565-1572
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• 2015

Porcine embryonic stem cells (pESCs) have become an advantageous experimental tool for developing therapeutic applications and producing transgenic animals. However, despite numerous reports of putative pESC lines, deriving validated pESC lines from embryos produced in vitro remains difficult. Here, we report that embryo aggregation was useful for deriving pESCs from in vitro-produced embryos. Blastocysts derived from embryo aggregation formed a larger number of colonies and maintained cell culture stability. Our derived cell lines demonstrated expression of pluripotent markers (alkaline phosphatase, Oct4, Sox2, and Nanog), an ability to form embryoid bodies, and the capacity to differentiate into the three germ layers. A cytogenetic analysis of these cells revealed that all lines derived from aggregated blastocysts had normal female and male karyotypes. These results demonstrate that embryo aggregation could be a useful technique to improve the efficiency of deriving ESCs from in vitro-fertilized pig embryos, studying early development, and deriving pluripotent ESCs in vitro in other mammals.