• Asian-Australasian Journal of Animal Sciences
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• 제29권8호
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• pp.1095-1101
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• 2016

Ginsenoside Rg1 is a natural compound with various efficacies and functions. It has beneficial effects on aging, diabetes, and immunity, as well as antioxidant and proliferative functions. However, its effect on porcine embryo development remains unknown. We investigated the effect of ginsenoside Rg1 on the in vitro development of preimplantation porcine embryos after parthenogenetic activation in high-oxygen conditions. Ginsenoside treatment did not affect cleavage or blastocyst formation rates, but did increase the total cell number and reduced the rate of apoptosis. In addition, it had no effect on the expression of four apoptosis-related genes (Bcl-2 homologous antagonist/killer, B-cell lymphoma-extra large, Caspase 3, and tumor protein p53) or two metabolism-related genes (mechanistic target of rapamycin, carnitine palmitoyltransferase 1B), but increased the expression of Glucose transporter 1 (GLUT1), indicating that it may increase glucose uptake. In summary, treatment with the appropriate concentration of ginsenoside Rg1 ($20{\mu}g/mL$) can increase glucose uptake, thereby improving the quality of embryos grown in high-oxygen conditions.

• 한국수정란이식학회지
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• 제13권2호
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• pp.107-115
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• 1998

The present study was carried out for the comparative study on the collection of bovine follicular oocytes by ultrasound-guided ovum pick-up(OPU) and slaughterhouse-derived (SHD) ovary aspiration and in vitro production of bovine embryos with the follicular oncytes in Korean native cows. Bovine follicular nocytes were observed with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use and the oocytes were collected with the aspiration equipment attached to the ultrasonograph. Bovine ovaries were collected and transported in phosphate buffered saline from the local slaughterhouse, the follicular oocytes were collected by the aspiration method. The collected follicular oocytes in good quality were matured, fertilized and cultured in the media. The total number of the visible follicles and the recovery rate of follicular oocytes were increased in ultrasonography following follicle stimulating hormone(FSH) treatment in Korean native cows. The mean recovery rate of oocytes was 66.2, 52.8 and 41.7% in the FSH-OPU, non-treatment-OPU and SHD ovaries, respectively. The mean number of recorved oocytes per cow were not significantly(P<0.05) different between the FSH-OPU(14.0$\pm$11.54) and SHD(17.1i6.21) groups, but the numbers in both groups were significantly(P<0.05) higher than the number in the non-treatment-OPU(3.7$\pm$1.57) group. The mean number of usable nocytes in Grade T /11 per ovary was 6.3, 4.8 and 1.3 in the cows of the SHD, FSH-OPU and non-treatment-OPU groups, respectively. The in vitro developmental rate to the blastocyst was not significantly different between the oocytes obtained via OPU(37.1%) and SHD(29.3%). Therefore, the ultrasound-guided OPU technique can be applied to the production of excellent embryos from the high-quality cows, and for the large scale production of in vitro bovine oocytes and embryos, the SHD ovary aspiration method is valuable.

• Clinical and Experimental Reproductive Medicine
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• 제28권4호
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• pp.265-270
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• 2001

Objective : The aim of this study was to evaluate the outcome the GnRH antagonist (Cetrotide) short protocol in controlled ovarian hyperstimulation comparing with GnRH agonist long protocol. Materials and Method: From July 2000 to November 2001, 26 patients, 28 cycles were performed in controlled ovarian hyperstimulation by GnRH antagonist and GnRH agonist. GnRH antagonist (Cetrotide) was administered in 12 patients (14 cycles, Group 1) and GnRH agonist (Lucrin, Sub Q, Group 2) in 14 patients (14 cycles). Ovulation induction was performed by hMG (Pergonal) in group 1, and by Combo (Metrodine HP + Pergonal) in group 2. We compared the fertilization rate, good quality embryo, and clinical pregnancy rate between the two groups. Student-t test and Chi-square were used to determine statistical significance. Statistical significance was defined as p<0.05. Results: Ovarian hyperstimulation syndrome did not occurred in which estradiol (E2) level was $3874{\pm}809\;pg/ml$ and the number of retrieved oocytes was $18.4{\pm}2.4$. The number of used gonadotropin ampules was significantly decreased in Group 1 (26.0 vs. 33.1, p<0.04). There were no significant difference in the number of preovulatory oocyte ($10.6{\pm}6.9$ vs. $10.0{\pm}6.1$), fertilization rate ($74.8{\pm}23.4$ vs. $72.2{\pm}21.8$), good quality embryo ($58.7{\pm}23.6$ vs. $38.7{\pm}36.6$), and embryo transfer ($4.3{\pm}1.6$ vs. $4.4{\pm}1.6$). Although the age of the group 1 was older than the group 2 (34.4 vs. 30.8), there was no significant difference in clinical pregnancy rate (50.0% vs. 57.1%). Conclusions: We suggest that GnRH antagonist was a safe, effective, and alternative method in the controlled ovarian hyperstimulation, especially in PCOD patients who will be develop the ovarian hyperstimulation syndrome.

• 한국수정란이식학회지
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• 제15권2호
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• pp.121-128
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• 2000

This study was conducted to enhance the efficiency of embryo transfer and embryos produced in vitro according to select good quality blastocyst among in vivo or in vitro produced embryos. After middle blastocys(MB) and late blastocyst(LB) stages embryos that developed in vitro for 7∼8 days were treated hypertonic solution(380 mOsm or 600 mOsm) and the rate of development to hatched blastocyst(HB) was examined. The results obtained were summarized as follows: The proportion of embryos developed to HB was higher than that of control regardless of hypertonic solution treatment. However, it was related to culture time in hypertonic solution The propotrion of embryos developed to HB of morphologically recovered embryos in 380 nOsm solution ws higher than that of morphologically shrinked embryos. However, treatment time was not significantly different the rates of HB development. Especially, the proportion of embryos developed to HB of morphologically recovered embryos of the 30-min-treated group was significantly higher than that of control (p<0.05). The results suggest that hypertonic solution treatment should enhance the efficiency for criterion of transferable blastocyst quality.

• Reproductive and Developmental Biology
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• 제30권1호
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• pp.21-26
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• 2006

본 연구는 체외성숙 배지에 첨가하는 PVP의 농도, EGF, cysteine 및 PVP의 단독 또는 혼합첨가가 한우 체외수정란의 체외발생에 미치는 영향을 검토하였다. 체외성숙 배지에 PVP의 첨가농도$(0.1{\sim}3.0%)$에 따른 분할율은 차이가 없었으나, 배반포 발달율은 0.5% PVP 첨가군이 가장 높았다(P<0.05). PVP, EGF 및 cysteine의 단독 및 혼합 첨가에 따른 분할율은 cysteine 단독첨가군이 높았으나(P<0.05), 배반포 발달율은 차이가 없었다. Inner cell mass 수는 대조군과 cysteine 첨가군이 PVP 첨가군에 비하여 유의하게 높았고(P<0.05), 총 세포수도 cysteine 첨가군에서 가장 높았다. 수정란이식 결과는 대조군, EGF, cysteine 및 EGF+cysteine 군의 임신율은 $46.1{\sim}63.6%$로서 비슷하였으나, PVP 첨가군은 10%로서 다른 군에 비하여 유의하게 낮았다(P<0.05). 본 연구 결과는 체외성숙 배지에 PVP의 첨가로 배 발생은 가능하지만, 세포의 품질에는 악영향을 미치는 것을 보여준다.

• Reproductive and Developmental Biology
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• 제37권4호
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• pp.281-287
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• 2013

In mammal, unfertilized oocytes remain in the oviduct or under in vitro culture, which is called "oocyte aging". This asynchrony negatively affects fertilization in pre- and post-implantation embryo development. Caffeine a phosphodiesterase inhibitor is known to rescue oocyte aging in several species. The objective of this study is to determine the cytoskeleton distribution in aged oocytes and the embryo developmental ability of aged oocytes in the present or absence of caffeine during maturation. Caffeine treatment increased the incidence of normal spindle assembly of aged oocytes (treatment, $67.57{\pm}4.11%$ aging, $44.61{\pm}6.4%$) and no significant differences compared to control group. Fluorescence values were compared using ROS (Reactive oxidation species) stain. Fluorescence values appear of control group intensity rate ($51.53.{\pm}3.80$), aging group ($68.10{\pm}5.54$) and treatment of caffeine ($45.04{\pm}2.98$). Aged oocytes that were derived from addition of caffeine to the IVM (in vitro maturation) medium had significantly increased 2-cell that developed to the blastocyst stage compared to the aging group. Blastocysts, derived from caffeine treatment group, significantly increased the total cell number compare aging ($90.44{\pm}10.18$ VS $67.88{\pm}7.72$). Apoptotic fragments of genomic DNA were measured in individual embryo using TUNEL assay. Blastocyst derived from caffeine treatment group decreased significantly the apoptotic index compared to blastocyst derived from aging group. In conclusion, we inferred that the caffeine treatment during oocyte aging can improve the developmental rate and quality in bovine embryos developing in vitro.

• Clinical and Experimental Reproductive Medicine
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• 제51권1호
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• pp.57-62
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• 2024

Objective: The purpose of this study was to use a mouse model to investigate the blastocyst formation rate in vitrified-warmed embryos derived from vitrified-warmed oocytes. Methods: Metaphase II oocytes obtained from BDF1 mice were vitrified and warmed, followed by fertilization with epididymal sperm. On day 3, a total of 176 embryos, at either the eight-cell or the morula stage, were vitrified-warmed (representing group 1). For group 2, 155 embryos at the same developmental stages were not vitrified, but rather were directly cultured until day 5. Finally, group 3 included day-5 blastocysts derived from fresh oocytes, which served as fresh controls. The primary outcome measured was the rate of blastocyst formation per day-3 embryo at the eight-cell or morula stage. Results: The rates of blastocyst formation per day-3 embryo were comparable between groups 1 and 2, at 64.5% and 69.7%, respectively (p>0.05). The formation rates of good-quality blastocysts (expanded, hatching, or hatched) were also similar for groups 1 and 2, at 35.5% and 43.2%, respectively (p>0.05). For the fresh oocytes (group 3), the blastocyst formation rate was 75.5%, which was similar to groups 1 and 2. However, the rate of good-quality blastocyst formation in group 3 was 57.3%, significantly exceeding those of group 1 (p=0.001) and group 2 (p=0.023). Conclusion: Regarding developmental potential to the blastocyst stage, vitrified-warmed day-3 embryos originating from vitrified-warmed oocytes demonstrated comparable results to non-vitrified embryos from similar oocytes. These findings indicate that day-3 embryos derived from vitrified-warmed oocytes can be effectively cryopreserved without incurring cellular damage.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.1-6
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• 1996

There are a number of problems during the process of culture in vitro on fertilization and embryo development compared to those on in vivo counterparts. And the platelet activating factor (PAF), which is found not only in mammalian spermatozoa but also preembryos, is implicated on reproductive process. To improve the environment of culture on in vitro fertilization and embryo development, coculture using salpingeal epithelial cells has been considered to accept the better result on pregnancy rate. This study was designed to determine if two different culture systems, coculture alone and PAF treated coculture, are positive or negative influence on process of in vitro fertilization and embryo culture in mouse. The cell cleavage rate reached to 2-4 cell stage at 24 hours of culture is 56.81% (50/88) and 48.21%(54/112) respectively, in PAF treated group which is added PAF on coculture and in coculture group. But the rate of cells cleavage was similar in both group after 48 hours of culture. The rate of unfertilization after insemination of oocytes was higher in coculture group(55..53%) than in PAF treated group(42.37%). And in assessment of undeveped embryos, the rate of equalized cell block was similar on both, coculture alone (35.3%)and PAF treated coculture(35.5%). while unequalized cell block was higher rate in PAF treated coculture(19.4%) than coculture alone (11.8%). But the rate of cytoplasmic degeneration of undeveloped embryos was significantly higher in PAF treated coculture than coculture alone. In conclusion, we have observed that PAF treated coculture is superior in the rates of in vitro fertilization and early embryo cell cleavage compared to those in coculture alone, but there is no difference on the rates of embryo develpments, cell degeneration, cell quality in both PAF treated coculture and coculture alone when the embryo cells were continuosly cultured for 48 hours or more.

• Clinical and Experimental Reproductive Medicine
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• 제43권4호
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• pp.221-227
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• 2016

Objective: The aim of this study was to evaluate the influence of maternal age on fertilization, embryo quality, and clinical pregnancy in patients undergoing intracytoplasmic sperm injection (ICSI) using testicular sperm from partners with azoospermia. Methods: A total of 416 ICSI cycles using testicular spermatozoa from partners with obstructive azoospermia (OA, n = 301) and non-obstructive azoospermia (NOA, n = 115) were analyzed. Female patients were divided into the following age groups: 27 to 31 years, 32 to 36 years, and 37 to 41 years. The rates of fertilization, high-quality embryos, clinical pregnancy, and delivery were compared across maternal age groups between the OA and NOA groups. Results: The rates of fertilization and high-quality embryos were not significantly different among the maternal age groups. Similarly, the clinical pregnancy and delivery rates were not significantly different. The fertilization rate was significantly higher in the OA group than in the NOA group (p< 0.05). Age-group analysis revealed that the fertilization and high-quality embryo rates were significantly different between the OA and NOA groups in patients aged 27 to 31 years old, but not for the other age groups. Although the clinical pregnancy and delivery rates differed between the OA and NOA groups across all age groups, significant differences were not observed. Conclusion: In couples using testicular sperm from male partners with azoospermia, pregnancy and delivery outcomes were not affected by maternal age. However, women older than 37 years using testicular sperm from partners with azoospermia should be advised of the increased incidence of pregnancy failure.

• 한국수정란이식학회지
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• 제19권2호
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• pp.133-145
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• 2004

난자의 체외성숙 및 체외배양에는 일반적으로 동물의 혈청을 사용하고 있다. 그러나, 채취한 소의 상태에 따라서 혈청의 질에 차이가 있어 실험데이터가 일정하지 않을 수 있고, 그것으로부터 바이러스, 세균, 마이코플라즈마 등에 오염될 가능성이 있다. 따라서, 본 실험에서는 완전 무 혈청 배양액에서 난자의 성숙, 배 발생율, 세포 수, 동결성을 검토하였다. 다음으로, 근래 혈청배지로 생산한 체외 배양 수정란은 과체중의 산자 생산, 초기 산자 사망률, caesarean section, dystocia 같은 발병으로 문제가 지적되고 있다. 그러나 무 혈청 배지로 생산한 체외 배양 수정란은 이러한 현상을 개선할 수 있다는 보고가 있어 본 실험에서는 완전 무혈청 배양액에서 생산된 동결란과 신선란을 젖소에 이식하여 그 결과를 검토하였다. 무혈청 배양액에는 에너지원과 세포성장인자가 첨가된 IVD101과 IVMD101, 대조군인 혈청 배양액에는 TCM199 + 10% FBS가 사용되었다. IVMD101에 의해 체외성숙이 이루어 졌고, 혈청이 첨가된 TCM199 + 10% FBS에서는 12.4% 배 발생율을 보인데 반해, 무혈청 배양액 IVD101과 IVMD101이 각 32.4%, 34.5%로 훨씬 높은 배 발생율을 보였다. 더욱이, 세포 수에 있어서도 무혈청 배양액에서 발생된 배반포의 세포수가 혈청이 첨가된 배양액에서 발생된 배반포의 세포수보다 월등하였으며, 이는 체내란과 비슷한 세포 수를 보이고 있다. 수정란의 내동성을 보면, 융해하고 24시간 배양기 정치 후에 IVD101과 IVMD101은 94.5%, 95.8%의 생존율을 보인 반면 TCM199 + 10% FBS는 52.5%에 불과하다. 융해하고 72시간후 탈출 배반포율이 IVD101과 IVMD101는 78.4%, 83.7%였는데 반해 TCM199 + 10% FBS는 32.0%였다. 마지막으로, 무혈청 배양액에서 발생한 동결란과 신선란을 젖소에 이식한 후에 임신율에 있어서는 유의적 차이는 없었지만, 무혈청 배지에서 생산된 수정란의 내동성의 탁월함이 증명되었다. 이러한 결과들로 종합해 보면, 무혈청 배양액 (IVD101, IVMD101)에서 발생된 수정란이 혈청 배양액에서 발생된 수정란보다 배발생율, 세포수, 동결성, 임신율에 있어서 우수함을 알 수 있었다. 이러한 무혈청 배양액은 배발생 과정의 연구뿐만 아니라 수정란 이식을 위한 고품질의 체외 수정란의 대량 생산, 복제, 형질전환 동물 생산 등에도 유익할 것으로 사료된다.