• Journal of Embryo Transfer
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• v.19 no.1
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• pp.1-10
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• 2004

These studies were carried out to improve the reproductive efficiency through embryos transfer of Hanwoo IVM/IVF embryos. Following routine IVM/IVF procedure, oocytes and zygotes were cultured far 40 to 44 h in CRlaa medium with BSA. Then 2 to 8-cell embryos were removed the cumulus cell and were cultured in CRlaa medium containing 10% fatal bovine serum and 2.5 mM taurine in 5% $O_2$ and 5% $CO_2$ at 38.5$^{\circ}C$. The fresh embryos of the morulae and blastocysts cultured for 6 to 9 days in vitro or the frozen-thawed embryos were transferred into recipients. The pregnancy rates of the blastocyst produced for 6, 7, 8, and 9 days in vitro culture were 59.4, 68.2, 66.0 and 100%, respectively. In the developmental stage, pregnacy rates of early blastocysts (61.1%), blastocysts(64.7%) and expanded blastocysts(69.5%) were higher than that of morulae stage(20.0%). The pregnancy rates according to the corpus luteum grades of A, B and C in recipients were 73.6, 62.9 and 50.0%, respectively. Effects of donor-recipients synchrony of after day 2, 1 and 0, before day 1 and 2 on the pregnancy rates were 35.7, 65.5, 72.6, 67.9 and 60.0%, respectively. Pregnancy rates of the body condition score of recipients $\leq$2(71.3%) were higher than those of $\geq$3.0 score(40.0%). The pregnancy rates according to the parity of recipients when embryo was transferred to cow(70.6%) was higher than in heifer(59.1%). The pregnancy rates according to hormone treatment before embryo transfer were 69.9% in hCG + GnRH administration group and 63.0% in control group. Fresh and frozen-thawed embryos on the pregnancy rates were 70.6 and 36.4%, respectively. Pregnancy rates in single and AI+single was 90.0% and 64.8%. Pregnancy rates in twin induction was better than in single. These results indicate that pregnancy rates after transfer were affected on the embryo ages, donor-recipient synchrony, body condition score of recipients, corpus luteum status, parity and hormone treatment to recipients.

• Clinical and Experimental Reproductive Medicine
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• v.39 no.2
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• pp.52-57
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• 2012

Objective: During IVF, non-transferred embryos are usually selected for cryopreservation on the basis of morphological criteria. This investigation evaluated an application for array comparative genomic hybridization (aCGH) in assessment of surplus embryos prior to cryopreservation. Methods: First-time IVF patients undergoing elective single embryo transfer and having at least one extra non-transferred embryo suitable for cryopreservation were offered enrollment in the study. Patients were randomized into two groups: Patients in group A (n=55) had embryos assessed first by morphology and then by aCGH, performed on cells obtained from trophectoderm biopsy on post-fertilization d5. Only euploid embryos were designated for cryopreservation. Patients in group B (n=48) had embryos assessed by morphology alone, with only good morphology embryos considered suitable for cryopreservation. Results: Among biopsied embryos in group A (n=425), euploidy was confirmed in 226 (53.1%). After fresh single embryo transfer, 64 (28.3%) surplus euploid embryos were cryopreserved for 51 patients (92.7%). In group B, 389 good morphology blastocysts were identified and a single top quality blastocyst was selected for fresh transfer. All group B patients (48/48) had at least one blastocyst remaining for cryopreservation. A total of 157 (40.4%) blastocysts were frozen in this group, a significantly larger proportion than was cryopreserved in group A (p=0.017, by chi-squared analysis). Conclusion: While aCGH and subsequent frozen embryo transfer are currently used to screen embryos, this is the first investigation to quantify the impact of aCGH specifically on embryo cryopreservation. Incorporation of aCGH screening significantly reduced the total number of cryopreserved blastocysts compared to when suitability for freezing was determined by morphology only. IVF patients should be counseled that the benefits of aCGH screening will likely come at the cost of sharply limiting the number of surplus embryos available for cryopreservation.

• Journal of Embryo Transfer
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• v.6 no.2
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• pp.37-46
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• 1991

A field trial was performed to evaluate the effects of hormone treatment on estrus induction, ovulation, embryo transfer and reproductive performance in post-weaning sows. This trial involved 61 mixed breed sows of varying parity on a commercial pig farm. Sows were allocated to one of five trials: control group involved 25 sows that were treated with a single intramuscular injection of 5 ml physiological saline, 6 sows received 1,500 IU PMSG on the day of weanning and 500 IU HCG at the onset of estrus in trial I, 7 sows received 750 IU PMSG on the day of weanning and 500 IU HCG at the onset of estrus in trial II, 5 sows were treated with the same as trial II on day 28 after weanning in trial III. and 18 sows were treated with 10 mg PGF$_2$$\alpha$ plus 2 mg estradiol benzoate on day 31 after weanning in trial IV. Ovarian responses were checked by laparotomy and ova were recovered by oviducal flushing between 40 and l00hrs after mating. Fertilized ova were transferred into the oviduts of recipient sows synchroni- zed. The results obtained were summarized as follows: 1. Percentages of sows detected in standing estrus following treatment were 86~100% among trial groups. The interval from treatment to standing estrus(6l.7$\pm$0.5lhrs) in lOmg PGF$_2$$\alpha$ and 2mg estradial henzoate treated group was significantly earlier than in other trial groups(P<0.05). 2. Average number of ovulations was 11.5~37.8 among trial groups. The ovulation rate in 1,500 IU PMSG and 500 IU RCG treated group (37.8$\pm$ 19.87) was significantly different from other trial groups(P<0.05). 3. Ova were recovered by oviducal flushing between 40~ l00hrs after mating and recovery rates of ova wore 91.4% between 40~59hrs. 4. Fertilized ova were transferred into the oviducts of 8 recipient sows synchronized with 7 to 17 ova per animal. Three of the recipients were pregnant and delivered 25 piglets. 5. Four of the donor sows in those embryo collection was not successful were pregnant following oviducal flushing and delivered 23 piglets. 6. Recurrence of estrus and farrowing performance of experimental sows were observed following the experiment was no difference among trial groups, respectively.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.179-183
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• 2016

Even though klotho deficiency in mice exhibits multiple aging-like phenotypes, studies using large animal models such as pigs, which have many similarities to humans, have been limited due to the absence of cell lines or animal models. The objective of this study was to generate homozygous klotho knockout porcine cell lines and cloned embryos. A CRISPR sgRNA specific for the klotho gene was designed and sgRNA (targeting exon 3 of klotho) and Cas9 RNPs were transfected into porcine fibroblasts. The transfected fibroblasts were then used for single cell colony formation and 9 single cell-derived colonies were established. In a T7 endonuclease I mutation assay, 5 colonies (#3, #4, #5, #7 and #9) were confirmed as mutated. These 5 colonies were subsequently analyzed by deep sequencing for determination of homozygous mutated colonies and 4 (#3, #4, #5 and #9) from 5 colonies contained homozygous modifications. Somatic cell nuclear transfer was performed to generate homozygous klotho knockout cloned embryos by using one homozygous mutation colony (#9); the cleavage and blastocyst formation rates were 72.0% and 8.3%, respectively. Two cloned embryos derived from a homozygous klotho knockout cell line (#9) were subjected to deep sequencing and they showed the same mutation pattern as the donor cell line. In conclusion, we produced homozygous klotho knockout porcine embryos cloned from genome-edited porcine fibroblasts.

• Journal of Embryo Transfer
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• v.11 no.3
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• pp.211-216
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• 1996

A series of experiments were designed to determine the feasibility of single dose of FSH as an effective superovulation method in rabbits. The superovulation treatments examined comparatively were as follows: 1) a single injection of 100 IU PMSG, 2) Multiple injections of 1 IU Super-OV of 2X daily for 3 days, 3) Multiple injections of 6.6 rng Folltropin-V 2X daily for 3 days, 4) Multiple injections of 13.3 rng Folltropn-V l$\times$ daily for 3 days, 5) Single injection of 40 mg Folltropin-V solved in 10% PVP, and 6) Single injection of 40 mg Folltropin-V dissolved in 25% PVP. The results obtained were as follows: The nurnher of ovulation points in the single injection of 40 mg Folltropin-V dissolved in 25% PVP and multiple injections of 6.6 mg Folltropin-V 2X daily for 3 days was averaged 44.6 and 39.2, respectively, which were significantly (P<0.05) more than in a single dose of PMSG or multiple doses of Super-OV. The number of total ova recovered in the single injection of 40 mg Folltropin-V dissolved in 25% PVP and multiple injections of 6.6 mg Folltropin-V 2X daily for 3 days was averaged 30.4 and 25.9, respectively, which were significantly (P<0.05) more than in the single dose of PMSG or multiple doses of Super-OV. A similar result was obtained for the number of fertilized ova recovered from the above six treatments. From the above results it was concluded that a single injection of 40 mg Folltropin-V dissolved in 25% PVP could be an effective and simple method for superovulation in rabbits.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.5 no.1
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• pp.94-107
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• 2005

Prenatal diagnosis (PND) such as amniocentesis or chorionic villi sampling has been widely used in order to prevent the birth of babies with defects especially in families with single gene disorderor chromosomal abnormalities. Preimplantation genetic diagnosis (PGD) has already become an alternative to traditional PND. Indications for PGD have expanded beyond those practices in PND (chromosomal abnormalities, single gene defects), such as late-onset diseases with genetic predisposition, and HLA typing for stem cell transplantation to affected sibling. After in vitro fertilization, the biopsied blastomere from the embryo is analyzed for single gene defect or chromosomal abnormality. The unaffected embryos are selected for transfer to the uterine cavity. Therefore, PGD has an advantage over PND as it can avoid the risk of pregnancy termination. In this review, PGD will be introduced and application of PGD in inborn error metabolic disorder will be discussed.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.269-274
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• 2003

The purpose of this study was to determine the effect of rbST treatment on progesterone concentration, volume of luteal tissue and pregnancy rate following embryo transfer. Recipient cows were assigned to control and rbST group, of which was given a single injection of rbST (500 mg. sc) at estrus detection. The concentration of progesterone was not significantly different between control and at 0, 3, 6 days after rbST treatment. However, the concentration of progesterone at 9, 12 days was significantly higher than in control group (4.6 and 6.8 vs. 3.9 and 4.5 ng/ml P4). The pregnancy rate after embryo transfer in rbST treatment was significantly higher than in control group (64.0 vs. 47.1 %; p<0.05). The results indicated that rbST treatment in recipient cows could be improved the efficiency of pregnancy rate after embryo transfer.

• Journal of Yeungnam Medical Science
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• v.37 no.1
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• pp.47-53
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• 2020

Background: This study was conducted to analyze clinical factors that can affect pregnancy rates in normal responders undergoing the freeze-all policy in in vitro fertilization. Methods: We evaluated 153 embryo transfer cycles in 89 infertile women with normal response to controlled ovarian stimulation (COS). After COS, all embryos were cultured to the blastocyst stage, and good quality blastocysts were vitrified for elective frozen-thawed embryo transfer (FET). Clinical variables associated with COS and the results of COS and culture, including the number of retrieved oocytes, fertilized oocytes, and frozen blastocysts were compared between the pregnant group and the non-pregnant group. Results: After a single cycle of COS for each patient, 52 patients became pregnant while 37 did not. Significant differences were observed in the number of matured oocytes, fertilized oocytes, frozen blastocysts, and transferred embryos. The number of frozen blastocysts in the pregnant group was almost twice that in the non-pregnant group (5.6±3.1 vs. 2.8±1.9, p<0.001). The area under the receiver operating characteristic curve for the 4 frozen blastocysts was 0.801 in the pregnant group. Conclusion: In the freeze-all policy, the number of matured oocytes, number of fertilized oocytes, and number of frozen blastocysts might be predictive factors for pregnancy.

• Journal of Embryo Transfer
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• v.11 no.1
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• pp.7-14
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• 1996

The object of this study was to evaluate the effect of uterine epithelial cells on development of Korean native cattle(KNC) oocytes fertilized in vitro. Qocytes were collected from ovaries of slaughtered Korean Native Cows and matured in TCM199 with granulosa cells supplemented with 10% FBS, 5$\mu$g/ml FSH, 10 JU/ml hCG, and 1$\mu$g/ml estradiol-17$\beta$ for 24 hrs. For co-culture of in vitro development of fertilized ova, oviductal epithelial cells (l$\times$l0˚cells /ml) obtained from slaughtered cow and uterine epithelial cells(1$\times$10˚cells /ml) flushed from the superovulated holstein on Day 7 were incubated in 39$^{\circ}C$, 5% $CO_2$, 95% air. Frozen-thawed KNC sperm was capacitated with BO(Brackett & Oliphant, 1975) medium supplemented with 10mM, 5mM-caffein. Matured oocytes were inseminated for 20 hrs. And then fertilized oocytes were washed with culture medium and transferred to oviductal epithelial cells for in vitro development and three days later a portion of embryos were transferred to uterine epithelial cells. Stastical methods of developmental rates on KNC-IVF oocytes was ANOVA-test. Developmental rates of KNC-IVF oocytes was significant higher(P<0.01) when co-cul-tured with uterine epithelial cells(25.2%) than oviductal epithelial cells. Blatocyst cul-tured for 7 to 9 days were frozen by automatic freezer with 1.4M glycerol-PBS. Survival rates of blastocyst was 40.0%. Fourteen frozen-thawed blastocysts were transferred to five holstein heifers on day 7 after natural estrus. Three recipients were observed twin and one recipient was single by ultra-sound systems on days 45 after embryo transfer.

• Journal of Plant Biology
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• v.32 no.4
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• pp.227-233
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• 1989

Single cells obtained from suspension culture of mature embryo-derived callus in wheat(Triticum aestivum L. cv Jang Kwang) were cultured to regenrated into the plantlet. Cell clusters and embryogenic calluses were efficiently developed from when the single cells clutured on the MS medium supplemented with 10${\mu}{\textrm}{m}$ 2,4-D. Upon transfer to hormone-free MS medium containing 10 mg/I AgNO3, embryogenic calluses gave rise to shoots, probably through somatic embryogenesis.