• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.26-26
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• 2003

Hyaluronic acid (HA)-binding sites have been shown the diagnostic potential fur assessment of sperm maturity, which is related to male fertility. This study was designed to evaluate chromosomal patterns in porcine embryos produced by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with non- or HA-binding sperm (HABS). For binding of sperm with HA, sperm incubated in 10 ${mu}ell$ drop containing HA (0.8 mg/ml)-agarose (0.8%) mixture for 15 min. IVF and ICSI with non- or HA-bound sperm examined with matured oocytes at 44 hr after in vitro maturation. Embryos were cultured in 50 ${mu}ell$ of NCSU 23 containing 0.5% BSA for 5 days and then in 50 ${mu}ell$ of NCSU 23 containing 10% FBS for 2 days. For the evaluation of chromosomal aneuploidies, chromosome 1 sub-metacentric specific probe was used in sperm and embryos by fluorescence in situ hybridization (FISH). The frequency of aneuploidy sperm for chromosome 1 was 6.25%. The significant differences following IVF and ICSI with non- or HA-bound sperm were not observed in blastocyst formation rates (18.6, 23.5, and 23.8%) and cell number (61.8 $\pm$ 12.5, 55.5 $\pm$ 7.3, and 59.3 $\pm$ 9.6). Moreover, the percentage of diploidy in 4-cell stage embryos was 57.1% (IVF), 68.8% (ICSI), and 76.3% (ICSI-HABS). These results suggest that HA-binding sites may be a material for selection of normal sperm for ICSI. Therefore HA selection of normal sperm may be reduce the loss to embryonic mortality prior to embryo transfer in pig.

• Reproductive and Developmental Biology
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• 제31권2호
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• pp.61-69
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• 2007

This study was designed to evaluate effects of BSA, PVA, gonadotropins and follicle shell during IVM of porcine oocytes and subsequent development to the blastocyst stage after IVF. Cumulus oocyte complexes (COCs) were cultured in TCM-199 media containing 4 mg/ml BSA and 1 mg/ml PVA during IVM for 44 hr. To compare the effect of gonadotropins on oocyte maturation, COCs were cultured with FSH+LH, FSH, LH and FSH-LH-free media during IVM. respectively. Also, different number of follicle shells (0, 2, 4 and 6) was used to examine whether the presence of follicle shell in culture medium affects oocyte maturation. The percentages of fertilization and blastocyst formation, respectively, were higher in the medium containing the PVA (49.0 and 17.9%) than those containing the BSA (40.0 and 12.2%). Significantly higher rates of Mil oocytes were in the presence of FSH+LH and FSH (88.6 and 85.1 %) compared to other treatments (64.0 and 53.4% at LH and FSH-LH-free media). Co-culture with inverted follicle shells in 2 ml maturation medium enhanced the developmental competence of porcine oocytes. In conclusion, PVA could be used as a macromolecules instead of BSA, and FSH and follicle shell played important roles in maturation of porcine oocytes.

• 한국수정란이식학회지
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• 제14권2호
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• pp.121-129
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• 1999

The objective of this study was to produce calves by transfer of embryos derived from slaughter house(SHD) and ultrasound-guided ovum pick-up (OPU). At 60 hrs after injection of 400 mg FSH dissolved in 25% polyvinylpyrrolidone(PVP) by single dose, ultrasound-guided follicular oocyte aspiration was ferformed. Day-7 and day-8 blastocysts produced by in vitro maturation (IVM), fertilization (IVF) and culture(IVC) of the oocytes derived from SHD and OPU were nonsurgically transferred into recipients. The results obtained were as follows. The cleavage rate and the development rate to blastocysts were not significantly (P<0.05) different between the oocytes obtained by SHD (72.9% vs. 34.1%) and OPU (75.9% vs. 38.4%). The oocyte recovery rate from the number of follicles by ultrasound-guided aspiration were not significantly (P<0.05) different between Holstein (61.7%) and Hanwoo (60.1%), but the rate of oocytes useful for IVF was significantly (P<0.05) higher in Hanwoo (69.3%) than Holstein (59.6%). The cleavage rate and the development rate to blastocysts was not significantly (P<0.05) different between Holstein (74.9% vs. 39.2%) and recipients on day 8 of estrus cycle resulted in 13 pregnancies (34.2%). One of them was sacrificed during gestation period due to mastitis and another was aborted spontaneous. The resulting 14 calves were morphologically normal at birth. Seventy eight fresh OPU-IVF embryos were transferred into 21 recipients on day 8 of estrus cycles, resulting in pregnancy of 12 recipients (41.4%). Two of them were sacrificed during gestation period due to mastitis and the other two were aborted. Nevertheless, the 11 OPU-calves have been born normally.

• 한국수정란이식학회지
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• 제12권3호
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• pp.277-282
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• 1997

The purpose of this experiment was to determine the effects of thiol compounds, $\beta$-mercaptoethanol($\beta$-ME) and cystearrone with buffalo rat liver cell(BRLC) co-culture on the development and intracellular glutathione(GSH) concentrations of bovine embryos produced by in vitro inaturation(IVM) and in vitro fertilization(IVF). Bovine IVM /IVF embryos developed to 2~8 cell stage were co-cultured with BRLC in GRlaa with or without thiol compounds. The developmental rate beyond morulae stage in CRlaa containing 0, 10,25 and 50$\pi$M $\beta$-ME with BRLG were 63.0, 74.0, 72.3 and 77.1%, respectively. And the developmental rate with 0, 25, 50 and 75$\pi$M cystearnine with BRLC were 69.6, 77.6, 81.0 and 76.8%, respectively. The developmental rate beyond morulae stage of GRlaa containing thiol compound with BRLG group was higher than that of control group. The intracellular GSH concentrations of blastocysts cultured for 5 days in GRlaa containing 0 and 50$\pi$M $\beta$-ME or cysteamine with BRLG were 81.2 and 86.4, 83.2 and 84.2pM, respectively. The intracellular GSH concentrations of blastocysts in GRlaa containing thiol compounds with BRLG was slightly higher than that of control group The cell numbers of blastocysts were not difference in all experimental groups. These results indicate that thiol compounds with BRLG co-culture was increased the percentage of developed into morulae and blastocysts, and intracellular GSII concentrations of blastocysts embryos.

• 대한한방부인과학회지
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• 제35권2호
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• pp.28-41
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• 2022

Objectives: The purpose of this study is to investigate the effectiveness of moxibustion in infertility with In Vitro Fertilization and Embryo Transfer (IVF-ET). Methods: We searched 8 databases (Embase, PubMed, CiNii, CNKI, OASIS, ScienceOn, KMBASE, KISS)to identify eligible studies published before 2021 Oct. We included randomized controlled clinical trials (RCTs) using moxibustion in infertility with IVF-ET. The methodological quality of each RCT was assessed by the Cochrane risk of bias tool. Results: Two RCT studies were eligible in our review. The overall risk of bias was evaluated as unclear. The meta-analysis of 2 trials indicated that favorable results for the use of moxibustion with IVF-ET. Conclusions: This systematic review and meta-analysis of clinical trials suggests that moxibustion with IVF-ET can effect on Infertility patients. However, because of studies included analysis was biased due to unclear risk of bias and unreliable study design, future RCT studies and additional Meta-Analysis are needed to judge the supplementary treatment role of moxibustion in infertility with IVF-ET.

• Clinical and Experimental Reproductive Medicine
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• 제26권1호
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• pp.117-121
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• 1999

Although anejaculation is a relatively uncommon occurrence in the general population, over 12,000 new cases are reported annually. Anejaculation may result from spinal cord injury, retroperitoneal lymph node dissection, diabetes mellitus, transverse myelitis, multiple sclerosis, or psychogenic disorders. At least 30% of men with this problem are or will be married and many will seek help to remedy their infertile state. The evolution of technique and instrumentation over the last 30 years has made electroejaculation an accessible and acceptable form of therapy. Recent successes in inducing ejaculation by means of rectal probe electrostimulation or vibratory stimulation combined with assisted reproductive techniques, such as zygote intrafallopian transfer (ZIFT), gamete intrafallopian transfer (GIFT), and in vitro fertilization (IVF), have provided these men means of producing their own biologic offspring. We have experienced a successful pregnancy with electroejaculation and in vitro fertilization in a infertile patient whose husband had an ejaculatory disturbance due to a spinal cord injury. So we report this case with a brief review of literatures.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.103-108
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• 1996

Through the previous studies(I,II), it was observed that human follicular fluid(HFF) was more effective than human fetal cord serum(HFCS) on promoting meiotic resumption of oocytes and improving embryonic development of mouse in vitro. On the basis of these results, we have gradually exchanged HFCS with HFF as protein supplement in human ART. This study was performed to investigate the efficiency of HFF on improving the pregnancy rate in ART. Oocytes were retrieved transvaginally from patients treated with pituitary suppression with GnRH-agonist and ovarian stimulation with human menopausal gonadotro-pin(HMG) and pure follicle stimulating hormone(FSH). Aspirated oocytes were rinsed and cultured in TCM-199 containing HFF, and the concentrations of HFF were adjusted to 10, 20, and 30% according to the use for insemination, embryo growth and embryo transfer, respectively. As possible as, we tried to do embryo transfer into fallopian tube to mimic the coincidence of the cell stage with the place of sojourn in vivo, so we performed various ART programs(IVF & ET; in vitro fertilization, ZIFT; zygote intra fallopian-tube transfer, ZIFT & ET) according to the tubal conditions of patients. On the while, intra cytoplasmic sperm injection(ICSI) was used to assist IVF of the patients who had shown poor standard IVF results by immunological or severe male factor. Of the 255 cycles of ART programs using HFF as protein supplement, 118 cycles were turn out to be succeeded in pregnancy(46.2%, per cycle, p<0.05), while 21 pregnancies were achieved in the 69 cycles using HFCS(30.4%). The 255 cycles using HFF were subdivided into cycles with the type of ART programs, and each pregnancy rate of the ART programs were 44.7% (IVF & ET, 76/170 cycles), 53.4%(ZIFT, 31/58 cycles) and 40.7% (ZIFT & ET, 11/27 cycles), respectively. In the 61 ICSI cycles using HFF, 28 cycles succeed in pregnancy(45.9%), while 7 pregnancies were obtained in the 17 ICSI cycles using HFCS. Also the ongoing pregnancy rate in the group using HFF(78.8%, 93/118 cycles) was higher than that in the group using HFCS(61.9%). Therefore, we found that the use of HFF as protein supplement was more suitable and effective than the use of HFCS to improve the pregnancy rate in ART.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.48-48
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• 2002

Chimerism has become an important tool for investigating fundamental aspects of early embryonic development and differentiation in mammals for producing transgenic animals. The objective of this study was to evaluate the developmental capacity of chimeric embryos reconstructed with parthenotes and IVF bovine embryos into empty zona pellucida. The MII oocytes were activated by two treatment groups [Group 1, 5 μM inomycin, 5min, ＋ 10 ㎍/㎖ cycloheximide (CHX)/5 ㎍/㎖ cytochalasin B (CCB), 3 h; Group 2, 5 μM ionomycin, 5 min ＋ 1.9 mM 6-dimetylaminopurine (6-DMAP), 3 h]. (omitted)

• Clinical and Experimental Reproductive Medicine
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• 제16권2호
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• pp.161-171
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• 1989

The development of 2-cell mouse embryos to the blastocyst stage in vitro has been used as quality control for the culture media and supplements employed for human in vitro fertilization and embryo transfer(IVF-ET). 2-cell mouse embryos were cultured to the blastocyst stage in SECM, Medium 199-Earle's, Ham's F-10 I , Ham's F-10 II , Hoppe & Pitts, MEM and $HT_6$. The protein supplements contained in media were bovine serum albumine, fetal bovine serum and human fetal cord serum. The results were as follows; 1. The successful development was 81.3% in Medium 199-Earle’s, 91.9% in Ham’s F-10 I and 97.1% in $HT_6$. 2. 2-cell mouse embryos developed properly in all supplements but the best development was particularly noted in $HT_6$ media when HFCS was supplied as protein supplement.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.49-53
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• 2012

The present study was conducted to examine the generation of reactive oxygen species (ROS) during micromanipulation procedures in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine enucleated oocytes were electrofused with donor cells, activated by a combination of Ca-ionophore and 6-dimethylaminopurine culture. Oocytes and embryos were stained in dichlorodihydrofluorescein diacetate or 3'-(p-hydroxyphenyl) fluorescein dye and the $H_2O_2$ or $^.OH$ radical levels were measured. $In$ $vitro$ fertilization (IVF) was performed for controls. The samples were examined with a fluorescent microscope, and fluorescence intensity was analyzed in each oocyte and embryo. The $H_2O_2$ and $^.OH$ radical levels of reconstituted oocytes were increased during manipulation (37.2~49.7 and 51.0~55.2 pixels, respectively) as compared to those of mature oocytes ($p$<0.05). During early $In$ $vitro$ culture, the ROS levels of SCNT embryos were significantly higher than those of IVF embryos ($p$<0.05). These results suggest that the cellular stress during micromanipulation procedures can generate the ROS in bovine SCNT embryos.