• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.301-309
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• 1997

본 연구에서는 체외수정, 난자내 정자 직접주입, 난자내 정자 두부 미두 주입 후의 핵과 미세소관의 변화를 관찰하였다. 핵과 미세소관의 움직임은 형광염색을 실시한 후 공초점주사현미경을 이용하여 관찰하였다. 체외수정에서 관찰된 바와 동일하게 정자를 난자에 직접주입 한 직후 정자 중편부에서 성상체가 형성되었고, 이 성상체에 의해 자성 웅성 전핵이 융합되는 것으로 관찰되었다. 그러나 난자내 정자를 직접주입하였을 경우 웅성전핵으로 발달하는 비율이 낮았다. 이는 주입된 정자가 원형질막과 perinuclear theca에 싸인 체 난자내로 들어가 난자내의 sperm nucleus decondensing factor와 정자 핵과의 반응이 억제되기 때문으로 생각된다. 정자 두부 만을 주입하였을 경우 성상체가 형성되지 않았지만 자성 웅성 전핵 사이 또는 그 주위에서 두터운 미세소관층이 관찰이 되었다. 따라서 소에 있어서는 정자의 중편부에 위치하여 microtubule organizing center (MTOC)의 역할을 하는 중심립 또는 중심체 없이도 모계에서 유래된 미세소관이 형성되어 이것이 전핵의 융합과 세포분열에 관여하는 것으로 생각된다. 정자의 미부 만을 주입하였을 경우 성상체가 형성이 되지 않았으며, 자성핵 사이에 형성된 미세소관과 떨어져서 관찰되었다. 따라서 주입된 정자의 꼬리는 미세소관형성과 관련이 없는 것으로 생각된다. 이러한 결과는 소에 있어서, 수정 시 정자로부터 유래되는 중심립 또는 중심체가 없이도 미세소관을 형성하여 미세소관에 의해 이후의 배발달이 정상적으로 일어남을 보여주고 있다.

• Clinical and Experimental Reproductive Medicine
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• 제40권4호
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• pp.169-173
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• 2013

Objective: To measure Irish opinion on a range of assisted human reproduction (AHR) treatments. Methods: A nationally representative sample of Irish adults (n=1,003) were anonymously sampled by telephone survey. Results: Most participants (77%) agreed that any fertility services offered internationally should also be available in Ireland, although only a small minority of the general Irish population had personal familiarity with AHR or infertility. This sample finds substantial agreement (63%) that the Government of Ireland should introduce legislation covering AHR. The range of support for gamete donation in Ireland ranged from 53% to 83%, depending on how donor privacy and disclosure policies are presented. For example, donation where the donor agrees to be contacted by the child born following donation, and anonymous donation where donor privacy is completely protected by law were supported by 68% and 66%, respectively. The least popular (53%) donor gamete treatment type appeared to be donation where the donor consents to be involved in the future life of any child born as a result of donor fertility treatment. Respondents in social class ABC1 (58%), age 18 to 24 (62%), age 25 to 34 (60%), or without children (61%) were more likely to favour this donor treatment policy in our sample. Conclusion: This is the first nationwide assessment of Irish public opinion on the advanced reproductive technologies since 2005. Access to a wide range of AHR treatment was supported by all subgroups studied. Public opinion concerning specific types of AHR treatment varied, yet general support for the need for national AHR legislation was reported by 63% of this national sample. Contemporary views on AHR remain largely consistent with the Commission for Assisted Human Reproduction recommendations from 2005, although further research is needed to clarify exactly how popular opinion on these issues has changed. It appears that legislation allowing for the full range of donation options (and not mandating disclosure of donor identity at a stipulated age) would better align with current Irish public opinion.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.147-152
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• 2020

Objective: The purpose of this study was to determine the effect of vaginal progesterone for luteal phase support (LPS) on the clinical pregnancy rate (CPR) in natural frozen embryo transfer (FET) cycles via a meta-analysis. Methods: We performed a meta-analysis of randomized controlled trials (RCTs) and retrospective studies that met our selection criteria. Four online databases (PubMed, Embase, Medline, and the Cochrane Library) were searched between January 2017 and May 2017. Studies were selected according to predefined inclusion criteria and meta-analyzed using R software version 2.14.2. The main outcome measure was CPR. Results: A total of 18 studies were reviewed and assessed for eligibility. One RCT (n = 435) and three retrospective studies (n = 3,033) met the selection criteria. In a meta-analysis of the selected studies, we found no significant difference in the CPR (odds ratio [OR], 0.96; 95% confidence interval [CI], 0.60-1.55) between the vaginal progesterone and control groups. An analysis of the two retrospective cohort studies that reported the live birth rate (LBR) following FET showed a significantly higher LBR in the vaginal progesterone group (OR, 1.72; 95% CI, 1.21-2.46). A subgroup meta-analysis of FET conducted 5 days after injection of human chorionic gonadotropin showed no significant differences between the two groups with regard to the CPR (OR, 1.18; 95% CI, 0.90-1.55) or miscarriage rate (OR, 0.73; 95% CI, 0.36-1.47). Conclusion: The results of this meta-analysis of the currently available literature suggest that LPS with vaginal progesterone in natural FET cycles does not improve the CPR.

• 동의생리병리학회지
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• 제33권2호
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• pp.141-150
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• 2019

Despite the development of assisted reproduction technologies (ART) including in vitro fertilization (IVF), the poor ovarian response and endometrial receptivity remains clinically a major unmet need. Although these problems are difficulties to solve in infertility treatment, there are no good therapeutic option yet. Traditional herbal remedies and acupuncture, therefore are being proposed as alternative treatment. Our group found that traditional herbal medicines such as Paeonia lactiflora L.(PL, 芍藥), Cyperus rotundus L.(CR, 香附子), and Perilla frutescens (PF, 紫蘇葉) could improve endometrial receptivity. In this study, we found out Yeosin-san (如神散) as an optimal herbal formula via combination of the previously established herbal medicines. Yeosin-san is a traditional Korean medical formula which was established by Ziming Jin (陳自明) and recorded in Furendaiquanliangfang (婦人大全良方) at first. The formula traditionally used for treating abnormal uterine bleeding and leukorrhea. It showed a highest effect on leukemia inhibitory factor (LIF) expression and on the adhesion between trophoblastic cells and endometrial cells. In addition, it has been shown that the Yeosin-san not only increases the endometrial receptivity to improve the embryo implantation but also enhances the ovary function by expressing the angiogenesis-related genes. Here we suggest that Yeosin-san could be a novel and effective candidate for treating female infertility.

• 한국수정란이식학회지
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• 제29권1호
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• pp.13-19
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• 2014

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in $38.5^{\circ}C$, 5% $CO_2$ incubator. For freezing, Day 7~9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to $-7^{\circ}C$, and the straw was seeded during 8 minutes-holding time, and was cooled to $-35^{\circ}C$ at the cooling rate of $0.3^{\circ}C/min$, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method ($56.86{\pm}26.53%$) were slightly higher than those by the slow-rate freezing method ($55.07{\pm}26.43%$) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were $72.65{\pm}18.3%$ and $79.06{\pm}17.8%$, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were $66.22{\pm}18.8%$ and $45.76{\pm}12.8%$, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.111-118
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• 1997

Intracytoplasmic sperm injection (ICSI) recently has been utilized widely as the most successful technique to overcome the unfertilization problem in cases of severe male infertility in couples who could not be treated by conventional IVF. Recently, indications of ICSI have been extended further and more fertilized oocytes become available. Thus, it is necessary to examine the efficiency of freezing the surplus embryos obtained from ICSI. We compared the survival rate and the future outcome of cryopreserved embryos obtained either after conventional IVF or ICSI during the same period. After ICSI or IVF, five best-quality embryos from each patient were transferred in the stimulation cycle and the surplus pronuclear (PN) stage oocytes or multicellular embryos were cryopreserved by slow freezing protocol with 1,2-propanediol (PROH) as a cryoprotectant. A total of 792 embryos from ICSI trial were thawed and 65.2% (516/792) survived. The survival rates of PN stage oocyte, multicellular embryo and PN + multicellular embryo were 63.5%, 68.2%, 64.0%, respectively. After 111 transfers, 34 pregnancies were achieved, corresponding to a clinical pregnancy rate of 30.6% per transfers. We thawed 1033 embryos from IVF trials and 57.5% (594/1033) survived. In IVF cycle, the survival rates of PN stage oocyte, multicellular embryo and PN + multicellular embryo were 58.2%, 65.2%, 40.2%, respectively. Thirty eight clinical pregnancies were established after 134 transfers, corresponding to a pregnancy rate of 28.4% per transfer. The cleavage rate of thawed PN stage oocytes from ICSI trial (61.3%) was significantly higher than those from conventional IVF (53.4%). The developmental rates of good embryo (${\geqq}$ grade II) in thawed PN stage oocytes obtained from conventional IVF and ICSI were 63% and 65%, respectively. We concluded that PN stage oocytes, multicellular embryos resulting from ICSI procedure can be successfully frozen/thawed with reasonable clinical pregnancy rates comparable to those of IVF.

• Clinical and Experimental Reproductive Medicine
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• 제40권3호
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• pp.122-125
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• 2013

Objective: The majority of embryo transfers (ETs) to date have been performed on day 3 to reduce the potential risk of developmental arrest of in vitro cultured embryos before ET. Development of sequential media has significantly improved culture conditions and allowed blastocyst transfer on day 5. While day 5 ET provides higher clinical pregnancy outcomes with reduced risks of multiple pregnancies, it still has potential risks of developmental arrest of IVF embryos. The aim of this study was to evaluate the clinical outcomes of day 4 ETs and compare the efficacy of day 4 ET with day 5 ET. Methods: From 2006 to 2009, a total of 747 fresh IVF-ET cycles were retrospectively analyzed (day 4, n=440 or and day 5, n=307). The cycles with any genetic factors were excluded. The rates of matured oocytes, fertilization, good embryos, and clinical pregnancy of the two groups were compared. The chi-square test and t-test were used for statistical analysis. Results: There were no significant differences between the two groups with respect to the mean age of the females and rates of matured oocytes. The pregnancy outcomes of day 4 ET (40.7%) were similar to those of day 5 ET (44.6%). The implantation rate of day 5 ET (24.2%) was significantly higher than that of day 4 ET (18.4%) (p=0.003). Conclusion: Day 4 ET can be chosen to avoid ET cancellation in day 5 ET resulting from suboptimal circumstances in the IVF laboratory, but the decremented quality of embryos for transfer and the decreased pregnancy rate must be taken into consideration.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.31-37
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• 2003

연구목적: 일반적으로 IVF-ET에서 가장 높은 임신율을 얻는 방법은 5 day ET (배반포기 배아 이식)이지만 장기간 배양이 적절하지 못한 경우에는 $2{\sim}4$일째에 ET를 실시하고 나서 $5{\sim}7$일째에 배반포기에 도달한 배아를 재이식 (SET)하여, SET의 효용성에 대하여 조사하고자 실시하였다. 연구재료 및 방법: 48주기의 환자에서 회수한 난자와 수정란은 10%와 20% hFF가 첨가한 DMEM에서 이식 직전까지 각각 공배양하였다. 채란 2일 (group I, day 2 ET), 5일째 이식 (group II, day 5 ET) 또는 재이식 (group III, SET; 2-5, 2-6, 2-7, 3-5, 4-7일)을 실시하면서 수정률, 할구분할률 및 임신율을 각각 비교하였다. 결과에 대한 통계 분석을 SAS (version 6.2)를 이용한 Duncan's Multiple Range Test를 이용하여 p값이 0.05 보다 작을 때 통계적으로 유의차가 있는 것으로 하였다. 결 과: 수정률은 group II (90.5%)가 다른 군에 비하여 높게 (p<0.05) 나타났다 (group I: 80.6%; group III: 82.9%). 할구분할률은 군간에 차이가 없었다 (수정란 당 $93.3{\sim}99.1%$). 임상적 임신율은 group II와 III (각각 58.3%)가 group I (33.3%) 보다 높게 나타났다. 그러나 처리군이 적어서 통계적인 차이는 없었다. 결 론: 배반포기 배아를 단독 이식하는 것이 임신율을 높일 수 있는 최선의 방법으로 나타났지만, 채란수가 적거나 수정률이 저조한 경우에는 $2{\sim}4$일째에 ET를 실시한 후 여분의 배아를 배반포기까지 배양한 다음 $5{\sim}7$일에 재이식 (SET)하면 blastocyst ET에서 나타날 수 있는 이식 자체의 실패를 방지할 수 있으면서 임신율을 높일 수 있는 이식 기법이 될 것이다.

• 한국수정란이식학회지
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• 제9권1호
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• pp.95-102
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• 1994

본 연구는 종모우의 선발방법으로 난포란을 이용하여 실험실내 정자의 수정능력을 직접 검정하여 평가코자 시도되었다. 즉 본 실험은 후대검정중에 있는 한우 후보종모우 15두의 동결융해정자의 수정능력을 평가하기 위하여 정액을 고장액(HIS)에 처리한 후 DM에서 6시간 그리고 소 난포액이 20% 첨가된 DM에서 4시간 전배양하여 수정능을 획득시켜 정자의 활력과 첨체 반응율을 조사하였고 전배양된 정자의 체내(토끼 난관) 또는 체외수정능력을 조사하기 위하여 FCS 15%, 발정암소혈정(CSS) 10%가 첨가된 mKRB에서 체외성숙된 한우난포란과 수정시켜 수정능력을 평가하였으며 인공수정에 의한 개체별 수태율과도 비교 검토한 바 다음과 같은 결론을 얻었다. 1. 한우 난포란의 체외성숙율은 BSA 첨가구 에서 43.8%, FCS 15% 첨가구에서 67.4%, CSS10% 첨가구에서 69.9%이었다. 2. 토끼 난관에서 체외수정율은 BSA 참가구에서 43.8%, FCS 15% 참가구 41.2% 및 CSS 10% 참가구 35.0% 이었다. 3. 후보종무우 15두의 정액을 HIS-DM으로 처리후 6시간 전배양하였을 때 정자의 활력지수는 9-32%였고 첨체반응율은 19-44% 이었으며 20% 난포액을 첨가하여 4시간 전배양 하였을 때 정자의 활력지수는 9-13% 이었고 첨체반응율은 20-43%로 개체간에 차이가 있었다. 4. 체외수정율은 6.6-85.7%였으며, 발정암소혈청(CSS) 10%가 첨가된 mKRB에서 성숙시킨 난포란이 FCS 15% 첨가된 mKRB에서 성숙시킨 난포란보다 다소 높았으나, 정자수정능획득방법간에는 차이가 없었 다. 5. 체외수정율에 있어서 전배양후 정자활력지수와는 부의 상관이 었으며, 첨체반응율과는 낮은 정의 상 관을 나타냈다. 6. 종모우의 수태율은 체외수정율, 정자활력지수 및 첨체반응율과 낮은 정의 상관관계를 나타냈다. 7. 종모우의 개체간 수태율 우열순위에서는 수정율순위와의 사이에 더욱 낮은 부의 상관관계를 보였다. 8. 이상의 연구결과 비록 후대정검중의 제한된 자료로 인하여 종모우 수태율과 체외수정율간에 유의적 인 상관관계는 없었으나, 연결 한우 수정율 평가에 대한 실험실내의 검정가능성을 찾을 수 있었다.

• 한국가축번식학회지
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• 제26권2호
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• pp.153-163
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• 2002

본 연구에서는 레트로 바이러스를 이용하여 형질전환 돼지를 생산하기 위한 기초 연구로써 유전자 도입 효율을 증진시키기 위해 sucrose와 poly-brene을 사용하여 그 효과를 조사하였다. 유전자(hGH) 전달체로는 vesicular stomatitis virus gly-coprotein pseudotyped retroviral vector (VSV-G)를 사용하였다. 체외에서 성숙된 돼지 난자는 매우 협소한 위란강내 공간을 가지며 따라서 위란강내 공간의 확장은 필수조건이다. sucrose 처리는 수정율및 배발달에는 영향을 미치지 않았으며 sucrose 처리를 통해 돼지난자는 위란강의 공간이 확장되어 충분한 양의 바이러스 벡터의 주입이 가능하였다. PCR 분석 결과 8.3 %의 유전자 도입율을 나타낸 무처리군에 비해 0.5, 1, 2, 3 % sucrose 처리군에서 39.3, 43.3, 35.7, 40.7 %의 높은 유전자 도입율을 나타내었다. 추가적으로 돼지난자는 sucrose 처리에 의해 다정자 침입의 억제 효과를 나타내었다. 일반적으로 레트로 바이러스를 이용한 세포에의 높은 유전자 도입 효과를 나타내는 것으로 알려진 polybrene을 바이러스와 공동 주입한 결과 0.5, 5, 50$\mu\textrm{g}$/$m\ell$의 농도에서 각각 56.5, 50.0, 57.1 %의 도입율을 나타냄으로써 바이러스 단독 주입군의 34.6%보다 높은 효율을 나타내었다. 그러나 50$\mu\textrm{g}$/$m\ell$의 농도를 주입하였을 경우, 분할율과 배반포발달율이 43.3 및 4.6%를 나타냄으로써 바이러스 단독주입군의 72.0, 17.3%보다 유의적으로 낮았다. 이상의 결과를 종합할 때, sucrose의 처리는 돼지 난자의 발달에 영향을 주지 않으며 유전자 도입효율을 증진시킬 수 있었으며, 추가적으로 다정자 침입억제효과를 보였다. 또한 polybrene의 사용은 낮은 농도에서 유전자 도입효율을 증진시켰다. 이러한 결과들은 형질 전환 돼지의 생산 가능성을 높이는데 이용될 수 있다고 사료된다.