• 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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• 제37권1호
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• pp.57-64
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• 2010

목 적: 배양 환경과 동결 기술이 발달함에 따라 동결 포배의 해동-이식의 빈도가 증가하고 있으며, 신선 주기와 마찬가지로 해동-이식 주기에서도 질 좋은 배아를 선별하는 것은 임신 성공 여부를 결정하는 아주 중요한 과제이다. 본 연구는 동결 당시 포배로의 발달 속도가 임신 결과에 미치는 영향을 알아보기 위하여, 수정 후 5일 및 6일째 동결한 포배의 해동-이식 후 임신율을 비교 분석하였다. 연구방법: 2006년 1월부터 12월까지 5일째 또는 6일째 동결한 포배를 해동하여 2007년 6월까지 융해 이식한 87명, 93주기를 대상으로 하였다. 동결법은 ethylene glycol과 DMSO를 이용한 유리화 동결법을 이용하였으며, 팽창 포배는 인위적인 수축을 시행 후 동결하였다. 해동 과정은 이식 전날 시행하여 15~18시간 배양액에서 배양 후 재팽창 여부를 확인하였다. 결 과: 5일째 동결한 포배를 해동-이식한 52주기와 6일째 동결한 포배를 해동-이식한 41주기에서 환자의 나이, 이식한 배아의 수, 해동 배아의 생존율 등 임신 결과에 영향을 미칠만한 요인들의 차이는 없었다. 그러나 생화학적 임신율, 임상적 임신율, 진행 임신율, 착상율 등은 5일째 동결한 포배를 해동-이식한 주기에서 높게 나타났다. 결 론: 5일째 동결한 포배를 해동-이식했을 때의 임신율은 6일째 동결한 포배를 해동-이식했을 때의 임신율보다 2배 이상 높았으며, 이는 신선 주기와 마찬가지로 해동-이식 주기에서도 동결 전 배아의 발달 속도의 차이를 임신 성공 예측의 중요한 지표로 사용할 수 있음을 시사한다.

• 한국수정란이식학회지
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• 제27권4호
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• pp.253-258
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• 2012

This study was carried out to effects of ethylene glycol concentration, sucrose and culture day of in vitro production embryo on slow-down freezing in Hanwoo. 6, 7, 8 and 9 day embryos produced in vitro were frozen using 1.8M EG+0.1M sucrose, 1.8M EG+0.5% BSA and 1.5M EG+0.1M sucrose media. Survivability was confirmed after frozen-thawed 24 and 48h and ICM, TE cell number were counted by Hoechst 33342 and PI staining after frozen-thawed 24h. As a result, 1.8M EG+0.1M sucrose group was most significantly (p<0.05) higher compared with the other treatment groups on survivability, TE and total cell number after frozen-thawed 24h ($94.2{\pm}2.6%$, $94.67{\pm}3.4$ and $129.67{\pm}5.5$). ICM number did not found significant (p<0.05) differences between the three treatment groups. in 6, 7, 8 and 9 day of embryos using three types of freezing media, frozen-thawed, 1.8M EG+0.1M sucrose groups with embryos cultured 8 day was significantly (p<0.05) highest survivability to $98.3{\pm}1.7%$ after frozen-thawed 24h. 1.5M EG+0.1 sucrose group with embryos cultured 9 day was significantly higher survivability than group of embryos cultured 8 day after frozen-thawed 24 and 48h. In conclusion, 1.8M EG+0.1M sucrose media is considered to be effective to cryopreservation of embryos cultured 8 and 9 day.

• Clinical and Experimental Reproductive Medicine
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• 제49권3호
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• pp.196-201
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• 2022

Objective: This prospective consecutive study investigated the variation in sperm DNA fragmentation (SDF) in multiple semen samples from patients with cancer. Methods: Eighty-one patients with various cancers underwent multiple semen collections on 3 consecutive days for sperm cryopreservation prior to cancer treatment. A commercial Halosperm kit was used to measure SDF. Within- and between-subject coefficients of variation were estimated via random-effects analysis of variance to assess the consistency of semen parameters and SDF. Intraclass correlation coefficients (ICCs) were calculated to assess the magnitude of the between-subject component of variance relative to the total variance. Results: The volume of semen in the day-2 and day-3 samples was significantly lower compared with the day-1 sample. Most parameters showed high ICC values, suggesting that within-subject fluctuations were small relative to the between-subject variability. The highest ICC values were identified for the SDF (ICC, 0.68; 95% confidence interval [CI], 0.45-0.84) and semen volume (ICC, 0.67; 95% CI, 0.45-0.84). Conclusion: Our findings showed that repeated ejaculates from patients with cancer had stable SDF levels.

• Clinical and Experimental Reproductive Medicine
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• 제27권3호
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• pp.283-289
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• 2000

Objective: The study was performed to compare the survival rate and the development of day 2 mouse embryos which had freezing procedures done. Methods: We used three different vitrification solutions (EFS, VS14, DPS) and a ultrarapid freezing solution (UFS) for cryopreservation of day 2 mouse embryo. Results: We tested toxicity by exposing embryos to vitirification solutions and a ultrarapid freezing solution. The survival rates are 100%, 97.8%, 95.6% and 100% (EFS, VS14, DPS and UFS). After cultured for 96 hours, hatching rates of each group are 93.5% (no freezing), 95.6% (EFS), 86.4% (VS14), 93.0% (DPS), and 93.0% (UFS). There is no significant differences among groups. The survival rates after thawing cryopreserved embryos are 80.2%, 91.7%, 69.5%, 0% and 91.8% (slow freezing, EFS, VS14, DPS and UFS). Also cultured for 96 hours, the hatching rates are 93.5% (no freezing), 84.1% (slow freezing), 93.9% (EFS), 48.5% (VS14) and 70.1% (UFS). Conclusion: The survival rates of vitrification in EFS solution and ultrarapid freezing are higher than slow freezing (p<0.05). The hatching rate of vitrification in EFS solution cultured for 96 hours is highest, so vitrification of day 2 mouse embryos in EFS solution considered as more effective for cryopreservation.

• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.325-333
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• 1997

The use of hormonal stimulation in human in vitro fertilization and embryo transfer (IVF-ET) leads to increased production of embryos for ET. So to avoid high pregnancies and to allow conception in future, unstimulated cycles, cryopreservation of spare embryos is desirable. One of the improvement of cryopreservation methods is vitrification. We cryopreserved mouse day 3 embryos by vitrification using the three different vitrification solution (EFS40, VS11 and VS3a). EFS40 solution is consisted of 40% (v/v) ethylene glycol, Ficol170 30% (w/v) and 0.5M sucrose and VS11 is 6.0M ethylene glycol and 1.8M glycerol. And VS3a is 6.5M glycerol and 6% (w/v) BSA (bovine serum albumin). First we tested the toxicity of three vitrification solution by exposure to these solution during 3 min. After washing by thawing solution, the survival rates of each groups are 95.5%, 90.9% and 84.4% (EFS40, VS11 and VS3a). High percentages of them developed to expanded blastocyst and hatching embryos in culture 48hrs 94.2%, 97.7%, 100% and 97.4% (no treatment group, EFS40, VS11 and VS3a). So there is no significant differences among the each group. Second, after thawing of vitirfied embryos, the survival rates of each groups are 96.8% (slow freeze), 94.1% (EFS40), 85.5% (VS11) and 80.0% (VS3a, P vs. no freeze or EFS40 is 0.01). Vitrified embryos exhibited a high rate of development in vitro after 48hrs culture. The percentages of each group to blastocyst and hatching embryos are 88.7% (no freeze), 91.8% (slow freeze), 93.4% (EFS40), 87.7% (VS11) and 73.0% (VS3a, P vs. other group is 0.01). The results suggest that there is no significant differences in exposure of various vitrification solution and day 3 mouse embryos can be vitrified in solution EFS40 and VS11 by simple procedure.

• 한국작물학회지
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• 제49권4호
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• pp.354-357
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• 2004

A simplified technique which cryoprotects zygotic embryos by encapsulation-dehydration was developed for the germplasm conservation of herbaceous peony (Paeonia lactiflora Pall.). The highest survival rate $(85\%)$ was obtained from embryos treated by encapsulation-dehydration. The zygotic embryos were precultured on MS medium containing 0.3mg/L $GA_3$ for 1 day. The precultured embryos were encapsulated in $3\%$ (w/v) alginate beads and immersed for 1 h in MS medium containing 2 M glycerol and 0.5 M sucrose. The encapsulated embryos were dehydrated for 5h by air drying prior to direct immersion in liquid nitrogen. This encapsulation-dehydration method appears to be a promising technique for germplasm cryopreservation of a herbaceous peony.

• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.52-55
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• 2017

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.

• Journal of Animal Science and Technology
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• 제55권5호
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• pp.417-425
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• 2013

동결 닭 PGCs의 생식계열 키메라를 이용한 생체에의 복원을 실용화 하기 위해서는, 닭 PGCs의 동결보존기술의 향상에 의해 동결 및 융해 후의 많은 생존세포를 확보 하는 것과, 생식계열 키메라의 제작효율을 높이는 것이 반드시 필요하다. 닭 PGCs는 배양 5.5일령의 닭 원시생식선으로부터 채취하고, ACS 방법에 의해서 순수 닭 PGCs를 분리했다. 닭 PGCs의 동결보존실험결과 다음의 4종류의 동결방법을 각각 비교 검토했다. 1. 플라스틱 스트로에 의한 완만동결법 (SF), 동결보호물질은 2M 에틸렌 글리콜 (EG), 2. 스트로에 의한 급속동결법 (RF), 8M EG + 7% PVP, 3. 동결용 Cryotube에 의한 SF, 2M EG, 4. 튜브에 의한 SF, 10% DMSO. 동결 및 융해 후의 PGCs의 생존율은 각각 76.4%, 70.6%, 80.5%, 78.1%로 관찰되었다.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.43-50
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• 2006

The goal of this research was to develop an efficient cryopreservation protocol for germplasms of yam (Diosorea batatas), that were cultivated in Korea. Comparative studies with four other cryogenic techniques and subsequent experiments for shoot regrowth were conducted. in vitro-grown shoot-apices of the D. batatas were successfully cryopreserved by encapsulation-dehydration. The maximum survival of shoot-apices could be achieved when the precultured (with 0.3 M of sucrose for one day) and encapsulated (with a 3%(w/v) Na-alginate solution) apices were dehydrated for $3.5{\sim}4\;h$ prior to direct immersion in LN (liquid nitrogen). The frequency of regrowth rate of cryopreserved apices was not decreased during 3-month storage period. The thawing method markedly affected survival of the cryopreserved apices, and thawing at $40^{\circ}C$ for 3 min produced the best results. When cryopreserved apices were post-cultured on the post-culture medium (MS), supplemented with $0.2mgl^{-1}$ of BA ($N_6$-benzyladenine) and $0.2mgl^{-1}$ of kinetin, they showed direct shooting without callusing.