• 대한한방부인과학회지
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• 제32권2호
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• pp.119-128
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• 2019

Objectives: The aim of this case is to report the effects of herbal medicine on a single woman patient with a low level of AMH (anti-$M{\ddot{u}}llerian$ Hormone) in progress of Oocyte Cryopreservation. Methods: A patient with a low level of AMH had symptom of secondary amenorrhea. For preparing oocyte cryopreservation after a long time of secondary amenorrhea, she was treated by twice a day herb medication for 10 months. And we observed the effects of treatments by improvement of symptoms and following up endometrium ultrasonography. After oocyte cryopreservation, for maintaining her menstruation, she was also treated by twice a day herb medication for two and a half months. Results: After treatments, symptom of amenorrhea was improved and the thickness of endometrium was increased as well as AMH in progress of oocyte cryopreservation. So 20 oocytes could be cryopreserved. Conclusions: This case shows that herbal medicine can be a concurrent method for a single woman patient with secondary amenorrhea in progress of oocyte cryopreservation.

• Clinical and Experimental Reproductive Medicine
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• 제47권4호
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• pp.306-311
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• 2020

Objective: The aim of this study was to determine whether co-administration of a gonadotropin-releasing hormone (GnRH) agonist and human chorionic gonadotropin (hCG) for final oocyte maturation improved mature oocyte cryopreservation outcomes in young women with decreased ovarian reserve (DOR) compared with hCG alone. Methods: Between January 2016 and August 2019, controlled ovarian stimulation (COS) cycles in women (aged ≤35 years, anti-Müllerian hormone [AMH] <1.2 ng/mL) who underwent elective oocyte cryopreservation for fertility preservation were retrospectively analyzed. Results: A total of 76 COS cycles were triggered with a GnRH agonist and hCG (the dual group) or hCG alone (the hCG group). The mean age and serum AMH levels were comparable between the two groups. The duration of stimulation, total dose of follicle-stimulating hormone used, and total number of oocytes retrieved were similar. However, the number of mature oocytes retrieved and the oocyte maturation rate were significantly higher in the dual group than in the hCG group (p=0.010 and p<0.001). After controlling for confounders, the dual-trigger method remained a significant factor related to the number of mature oocytes retrieved (p=0.016). Conclusion: We showed improved mature oocyte collection and maturation rate with the dual triggering of oocyte maturation in young women with DOR. A dual trigger appears to be more beneficial than hCG alone in terms of mature oocyte cryopreservation for young women with DOR.

• 한국발생생물학회지:발생과생식
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• 제18권2호
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• pp.117-125
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• 2014

Vitrification method is widely used in oocyte cryopreservation for IVF but the birth rates are lower than that of the fresh oocyte. One of the known main reasons is structural instability of meiotic spindle and chromosome systems of mature oocyte. To get the best way for keeping competence of matured oocytes, we studied the best conditions for vitrification focused on equilibration times. The mature oocytes were underwent vitrification with current popular method and analyzed the survival rates, microtubule stability and DNA integrity. The survival rates of recovered oocyte are almost same between groups and are more than 93%. The structural configuration of meiotic spindle was well kept in 10 min equilibration group and the stability rate was almost same with that of control. The chromosomal breakdown was observed in all experimental groups, but the chromosomal stability was higher in 10 min equilibration group than the other groups. The 10 min equilibration group showed best condition compared with the other groups. Based on these results, the equilibration time is one of the key factors in successful keeping for competence of mature oocyte. Although, more fine analysis about the effects of physical stress on oocyte during vitrification is needed to define the optimal condition, it is suggested that the optimal equilibration time to get competent oocyte in mouse is 10 min. Information acquired this study may provide insight into intracellular structural events occurring in human oocytes after vitrification and application for cryopreservation of human oocyte.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.325-330
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• 1994

The study has been carried out in order to evaluate the effects of embryonic stage, and cryopreservation method on the rates of viability and development of the cryopreserved mouse early embryos. The results were as following:In the treatment steps of cryoprotectant, for the fertilized oocyte with pronucleus(PN), 2-step was better than the others. And for the other embryos, 4-step was better than 2- or 3-step. In respect to the embryonic stage, as the embryos developed from fertilized oocytes to 8-cell embryos, the rates of viability and development were increased higher. Therefore, 8-cell embryo was better stage than the others. In respect to the kind of cryoprotectants, PROH was better than DMSO for the fertilized oocyte, as a cryoprotectant. DMSO, for the 2-cell embryos and PROH and DMSO for the 4- and 8-cell embryos were suitable for cryopreservation.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2003년도 제44차 춘계학술대회
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• pp.57-57
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• 2003

• 한국수정란이식학회지
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• 제16권3호
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• pp.239-243
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• 2001

Selection of oocyte cryopreservation method is a prerequisite factor for developing an effective bank system. Compared with slow freezing method, the vitrification has various advantages such as avoiding intracellular ice crustal formation. In our previous, we attempted to employ a vitrification method using ethylene glycol and an electron microscope grid for cryopreservation of mouse oocytes. However, A high incidence of spindle and chromosome abnormalities was detected in thawed oocytes after vitrification. We examined whether the addition of a cystoskeleton stabilizer Taxol $^{TM}$, to the vitrification solution could promote the post-thawed survival and subsequent development of stored oocytes. More oocytes developed to the 4-cell (44.7% vs. 69.7%), 8-cell (31.8% vs. 64.2%), morula (24.7% vs. 54.3%), and blastocyst (20.3% vs. 49.2%) stages after the addition of Taxol$^{TM}$ to the cryoprotectant than after no addition. 21 and 26 mouse pups were born after transfer of blastocyst derived from oocytes vitrified without and with Taxol. The addition of Taxol to vitrification solution greatly promoted post-thaw preimplantation development of ICR morose oocytes.tes.

• 한국수정란이식학회지
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• 제30권4호
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• pp.277-282
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• 2015

Cryopreservation affects osmotic tolerance and intracellular ion concentration through changes in expression levels of water and ion channels. Control of these changes is important for cell survival after cryopreservation. Relatively little is known about changes in $K^+$ channel expression compared to water channel expression. This study was performed to investigate changes in TASK-2 channel (KCNK5: potassium channel, subfamily K, member 5), a member of two-pore domain $K^+$ channel family, in cryopreserved mouse ovaries. Cryopreservation increased TASK-2 mRNA expression in mouse ovaries. In addition, TASK-2 protein expression was upregulated in vitrified and slowly frozen ovaries. TASK-2 protein was expressed in all area of granulosa cells that surround the oocyte within the follicle, except nucleus. Viability of cells overexpressed with TASK-2 was higher than that of vector-transfected cells. Our results found that TASK-2 expression was increased by cryopreservation and overexpression of TASK-2 decreased cryopreservation-induced cell death. These results suggest that TASK-2 upregulation might reduce cryodamage.

• 한국수정란이식학회지
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• 제16권3호
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• pp.245-250
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• 2001

This study was performed to evaluate whether vitrification method using ethyle glycol and eletron microscopic (EM) grid could be used far the cryopreservation of human oocytes in ART program. Surplus oocytes were obtained from consented IVF patients. These surplus human oocytes were frozen with our vitrification method, Oocytes were exposed to 1.5M ethylene glycol (EG) in DPBS far 2,5 minutes, followed by 5.5M EG plus 1.0M Sucrose in DPBS for 20 seconds. Then oocytes were transferred onto the EM grid and the grid was plunged into LN2 for storage. For thawing, oocytes containing EM grid were sequentially transferred in 1.0M, 0.5M, 0.25M, 0.125M and 0 M sucrose in DPBS solution at the intervals of 2.5 minutes. Thawed and survived oocytes were provided for ICSI. Embryos from vitrified oocytes were transferred to uterus of the patient on 4 to 5 days after ovulation in natural cycles of on 15 to 17 day of hormone replacement cycles. A total of 370 oocytes from 26 patients were thawed and 159 (43.0%) of them survived. One hundred thirty four oocytes (84.3%) were fertilized normally and 126 pre-embryos were transferred to 26 patients, resulting in 5 clinical pregnancies. The pregnancy rate per transfer was 19.2% and implantation rate was 4.0%. Among the five pregnant, 4 patients delivered 4 healthy babies and the one patient was 32-week ongoing pregnancy. From this results, vitrification using ethylene glycol as cryoprotectant and EM grid is a rapid and simple method that can be effectively applied for the cryopreservation of human oocytes in ART program.

• Clinical and Experimental Reproductive Medicine
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• 제37권4호
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• pp.267-291
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• 2010

최근 동결기술이 발달하면서 다양한 목적에 따라 초기 발생단계, 특히 수정 전후의 난자나 수정란의 생명을 연장하는 것이 가능해졌다. 이러한 난자나 수정란의 보존기술은 인간의 수정능력을 배가시키거나 임신조절에서 응용되고 있으며, 동물에서는 우수한 유전자원의 보존과 운영, 저렴한 국제간 운송수단, 그리고 생식보조기술과 유전공학 등의 연구에 필요한 생식세포의 공급하는 데서도 중요하게 활용되고 있다. 최근 개발된 완만동결과 유리화 동결방법은 난자와 수정란을 장기간 동결하여 보존하는데 활용하는 주요 기술이다. 이러한 방법들은 각각 장점과 단점을 가지고 있지만, 상당한 수준의 효율성이 입증되어 실용화되어 있는 실정이다. 무엇보다도 유리화 방법은 완만동결 방법보다 13년이나 늦게 개발되었으나 보다 우수한 기술로 인정을 받고 있다. 비록 유리화 동결은 아직 대한 상반된 의견과 오염문제가 있지만 인간과 동물의 생식보조기술로 활용되는 빈도가 점차 많아지고 있는 실정이다. 따라서 본 원고에서는 먼저 난자와 수정란의 동결보존에 대한 기초적인 기술에 대해서 고찰한 다음, 유리화 동결에 관 한 최근의 연구동향에 대해서 종합적으로 검토하고자 한다.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2006년도 The 5th Biannual Meeting of Pacific Rim Society for Fertility and Sterility
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• pp.26-31
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• 2006