• Journal of Embryo Transfer
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• v.31 no.3
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• pp.215-219
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• 2016

Value of excellent breeding animals is important in livestock industry, but their economic life time is limited. And, many countries have been trying procuration of genetic resource in good animals. Therefore, this study was conducted to determine embryo production and to test efficiency of embryo transfer via non-surgical artificial insemination (AI) in different breed of superior sows. A total of 17 sows were used in this experiment (Duroc, n=10; Landrace, n=4; Yorkshire, n=3). The sows were artificially inseminated by semen of same breed boars. After 4 or 5 days following the AI, the embryos were obtained from the sows and then transferred to Landrace and Yorkshire recipients (n=3, respectively) by non-surgical method. The corpora lutea tended to be increased in Yorkshire and Landrace than Duroc(28 and 26 vs. 17, respectively). The recovery of embryo was 78.8% in Landrace, 65.4% in Duroc and 51.4% in Yorkshire. Duroc showed lower morulaes and early blastocyst embryos than 2, 4, 8 and 16 cell. The morula in Yorkshire was higher (P<0.05) than that of Duroc (4.7 vs. 3.4). Similarly, the morulaes and early blastocyst embryos presented greater (P<0.05) in Landrace compared with other breed sows. The recipient sows were pregnant in a Landrace only. This reason may be due to little embryos inserted in the recipients. In addition, pregnancy results were limited because of the little sows. In conclusion, ovulated ovum in sows can be affected by different breed. Also, further study needed pregnant test by using the many embryo in each breed.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.5
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• pp.621-628
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• 2012

The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.9
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• pp.1270-1275
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• 2014

An experiment was conducted to evaluate the efficacy of porcine follicle stimulating hormone (pFSH) dosage based on body weight (BW) on ovarian responses of crossbred does. Thirty donor does were divided into 3 groups getting pFSH dosages of 3, 5, and 8 mg pFSH per kg BW, respectively, and were named as pFSH-3, pFSH-5 and pFSH-8, respectively. Estrus was synchronized by inserting a controlled internal drug release (CIDR) device and a single injection of prostaglandin $F2{\alpha}$ ($PGF2{\alpha}$). The pFSH treatments were administered twice a day through 6 decreasing dosages (25, 25, 15, 15, 10, and 10% of total pFSH amount; decreasing daily). Ovarian responses were evaluated on Day 7 after CIDR removal. After CIDR removal, estrus was observed 3 times in a day and pFSH treatments were initiated at 2 days before the CIDR removal. All does in pFSH-5 and pFSH-8 showed estrus signs while half of the does in pFSH-3 showed estrus signs. No differences (p>0.05) were observed on the corpus luteum and total ovarian stimulation among the treatment groups, while total and transferable embryos were higher (p<0.05) in pFSH-5 (7.00 and 6.71) than pFSH-3 (3.00 and 2.80) and pFSH-8 (2.00 and 1.50), respectively. In conclusion, 5 mg pFSH per kg BW dosage gave a higher number of embryos than 3 and 8 mg pFSH per kg BW dosages. The results indicated that the dosage of pFSH based on BW is an important consideration for superovulation in goats.

• Journal of Embryo Transfer
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• v.15 no.2
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• pp.129-136
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• 2000

This study was carried out to investigate the effect of biological factors on the in vitro production(IVP) of bovine oocytes for development of simple culture methods and medium. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol and this study was examined if there were necessary to co-culture, media change, media type and embryo density. This results were as follows: 1. The development rate according to co-culture with cumulus cells and non co-culture as drop culture was not significantly different in cleavage (68.9 vs 71.7%), 8-cell stage (41.2 vs 44.1%) and blastocyst stage (12.2 vs 13.8%), respectively (p<0.05) 2. The blastocyst development rates in YS and CRIaa were higher than that in TCM199 (12.4, 10.4$ vs 3.7%), but the cleavage (69.0, 77.8 and 61.0%) and 8-cell stage (31.7, 37.0 and 35.7%) development accoring to YS, TCM199 and CRIaa ws not significantly different, respectively (p<0.05). 3. There was no significantly different in cleavage (62.6, 59.5 and 61.2%), 8-cell(34.7, 37.9 and 34.0%) and blastocyst (9.5, 11.6 and 12.8%) development among medium change time as control, Group I and Group II, respectively (p<0.05). 4. Blastocyst formation of 8-cell stage according to embryo density was not significantly different in 1, 10 and 25 embryos (27.3, 22.5 and 34.0%), respectively (p<0.05). These results indicated that simple culture system could reduce bovine IVP embryos as drop culture as non co-culture system, high density embryo (25 embryos/50 $\mu$1 drop). YS defined medium and no medium change for whole culture period, although other biological factors need to examine in order to produce efficient IVP bovine embryos.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.1
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• pp.50-56
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• 2019

These studies were conducted to evaluate developmental competence of follicular oocyte collected from the ovaries of Hanwoo cows with the high offspring meat quality (1++ and 1+ grade). Cumulus oocyte complexes from individual cows were matured, fertilized and cultured using protocols of in-vitro maturation (IVM), invitro fertilization (IVF) and in-vitro culture (IVC). The rates of blastocyst development from Hanwoo cows with the offspring meat quality grades of 1++ and 1+ were 18.6 and 21.2%, respectively. The rates of blastocyst development were 26.3, 20.7, 20.7, 17.2 and 31.2% from Hanwoo cows with the meat quality grades of 1++, 1+, 1, 2 and 3, respectively. Fiftyseven transferable embryos were recovered from 11 Hanwoo donor cows (5.2/head) with the high offspring meat quality grades of 1++ and 1+ in vivo, and the pregnancy rate after embryo transfer was 61.1%. In conclusion, these results suggest that in vitro embryo production from the ovaries of cows with the high meat quality grades using individual culture system can be used an efficient method for livestock improvement. In addition, for the successful industrialization of embryo transfer, conception rate should be improved.

• Proceedings of the KSAR Conference
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• 2009.06a
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• pp.20-21
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• 2009

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 1998.05a
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• pp.12-28
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• 1998

The technology of creating transgenic animals has a potential value in improving productivity and disease resistance of animals, gene therapy, drug pharming and production of model animals for certain diseases. Up to date, fairly low success rate of production of transgenic animals and a pronounced variability with respect to the expression of transgenes have been much observed. The mechanisms how to integrate the injected genes with a certain part of the genomes are unknown yet. Many techniques in gene transfer, beside microinjection, have been introduced and explored thus to improve the production efficiency of transgenic animals. In this article, the methods and efficiency of gene-transfer techniques, the detection and preselection of transgenes in embryos by PCR- and GFP-screenings and cloning of preselected transgenic embryos by nuclear transplantation are described and discussed. Some experimental results showed that the early screening and selection of integration of the injected gene with embryonic genome by polymerase chain reaction(PCR) and green fluorecence protein(GFP) were promising methods. Further, the application of nuclear transplantation technology to cloning and multiplication of the positively integrated genes in the cleaving embryos and embryonic cells will be beneficially used for the mass production of transgenic embryos and consequently improving the production efficiency in transgenic animals.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.169-175
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• 2013

Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{\mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{\mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.191-198
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• 2006

The objectives of this study was to produce valuable offsprings of Hanwoo and Jeju black cattle using in vivo embryo production and embryo transfer techniques during 5 years ($2001{\sim}2005$) in Jeju. Two hundred and eighty six Hanwoo and sixty nine Jeju black cattles, at random stages of the estrous cycle, received a CIDR. Seven days later, the animals were superovulated with a total of 400 mg pFSH ($Folltropin^{(R)}-V$) administered twice daily in constant doses (each 50 mg) over 4 days. On the 6th administration of FSH, CIDR was withdrawn and 25 mg $PGF_{2{\alpha}}$ was administered. Cows were artificially inseminated thrice after estrous detection at 12 hr intervals. The cows received $250{\mu}g$ GnRH at the time of 2nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Recipients were synchronized with donors by insertion of a CIDR for 7 days and administration of 25 mg $PGF_{2{\alpha}}$ at the time of CIDR removal. The collected embryos were transferred to 1,219 recipients by 6 transfer persons. The mean numbers of total ova and transferable embryos from Hanwoo and Jeju black cattle donors were 7.4 and 4.7, respectively The number of transferable embryos differed between Hanwoo (5.0) and Jeju black cattle (3.5, p<0.05), while that of total ova did not differ. Repeated superovulation treatments decreased (p<0.05) the ratio of numbers of the flushed animals vs. superovulated animals in Jeju black cattle, and the numbers of total ova and transferable embryos in Hanwoo. More transferable embryos were collected at summer (5.6) than winter (2.9, p<0.01). The mean pregnancy rate was 40%. The pregnancy.ate was affected by transfer year (2001<2004, p<0.05) and transfer person ($33.0{\sim}41.9%$, p<0.01), while not by donor (embryo) breed. These results showed that in vivo embryo preduction and embryo transfer techniques using CIDR for Hanwoo and Jeju black cattle donors as well as recipient, regardless of their estrous cycle, may enable a stable embryo production and recipient preparation.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.4
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• pp.253-273
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• 2007

Objective: This study was to find ways to let a manager or superintendent rationally and consistently inspect as well as let a embryologist precisely record [The Book of Supernumerary Embryo Preservation] and [The Book of Supernumerary Embryo Donation]. Methods: Based on the data collected between 1994 and 2004 in Clinic 44 (Maria Fertility Hospital), [The Present State about Production and Use of Embryos], [The Preservation of Supernumerary Embryos and Their Thaw State], [The Present State about Thaw and Use of Frozen Embryos], [The Present State about Donation and Charge of Frozen Embryos], [The Book about Frozen Embryo Discard], and [The Summarization Book about Management and Use of Frozen Embryos] were designed and recorded. Results: The production, use, preservation, discard and donation quantity of human embryos, the use and discard quantity of thawed embryos, and the cumulative embryo preservation quantity could be totalized in [The Present State about Production and Use of Embryos in Clinic 44]. Also, [The Preservation of Supernumerary Embryos and Their Thaw State in Clinic 44] supported "the supernumerary embryo preservation quantity" etc. In addition, [The Present State about Thaw and Use of Frozen Embryos in Clinic 44] or [The Book about Frozen Embryo Discard in Clinic 44] supported "the use and discard quantity of thawed embryos" etc. Moreover, "The embryo donation quantity" could be totalized in [The Present State about Donation and Charge of Frozen Embryos in Clinic 44]. Finally, [The Summarization Book about Management and Use of Frozen Embryos in Clinic 44] could be used for rational and consistent management or inspection. Conclusion: The present results suggest that the documents not only be standard data to record [The Book about Supernumerary Embryo Preservation in Clinic] and [The Book about Supernumerary Embryo Donation in Clinic] but can also be preserved as treatment references.