• Clinical and Experimental Reproductive Medicine
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• 제37권1호
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• pp.65-72
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• 2010

목 적: 본 연구는 체외수정술에서 배양 3일째 2개의 난할단계 배아 (2ET)와 3개의 난할단계 배아 (3ET)를 이식했을 때 각각의 임신성적을 비교하기 위하여 시행하였다. 연구방법: 2007년 1월부터 2009년 6월까지 한 명의 불임전문 의사에게 시술받은 2ET군 100명을 환자의 나이와 체외수정 주기의 특성을 고려하여 3ET군 100명과 비교한 후향적 환자군-대조군 연구를 시행하였으며, 두 군 모두 양질의 배아만을 이식하였다. 각각의 임신율, 착상율, 다태임신율을 비교하였다. 결 과: 환자의 특성, 체외수정 주기 및 배아의 특징은 두 군 간에 차이를 보이지 않았다. 2ET군과 3ET군 모두 비슷한 임신성적을 보여주었다; 착상율 (41.0% vs. 35.3%), 임신율 (58.0% vs. 60.0%), 임상적임신율 (55.0% vs. 59.0%), 진행임신율 (51.0% vs. 55.0%). 두 군 모두 높은 다태임신율을 보여주었으나, 3ET군에서 유의하게 더 높은 전체 다태임신율과 삼태임신율을 보여주었다 (30.9% vs. 50.8%, p=0.031; 1.8% vs. 11.9%, p=0.036). 결 론: 나이가 ��고 양질의 배아를 가진 좋은 예후를 예측할 수 있는 여성에서 체외수정술 시행 시 배양 3일째 2개의 배아를 이식하더라도 3개의 배아를 이식했을 때와 비슷한 임신성적을 얻을 수 있고, 다태임신 (특히, 삼태임신)을 줄일 수 있다.

• 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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• 제40권2호
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• pp.90-94
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• 2013

Objective: To evaluate the efficacy of earlier oocyte retrieval in IVF patients with a premature LH surge on hCG day. Methods: One hundred forty IVF patients (164 cycles) with premature LH surge on hCG day were included, retrospectively. We divided them into 2 study groups: LH surge with timed ovum pick-up (OPU) 36 hours after hCG injection (group B, 129 premature cycles), and LH surge with earlier OPU within 36 hours after hCG injection (group C, 35 cycles). Control groups were tubal factor infertility without premature LH surge (group A, 143 cycles). Results: The mean age (year) was statistically higher in group C than in groups A or B ($38.2{\pm}5.4$ vs. $36.2{\pm}4.2$ vs. $36.8{\pm}4.9$, respectively; p=0.012). The serum LH levels (mIU/mL) on hCG day were significantly higher in group B and C than in group A ($22.7{\pm}14.9$ vs. $30.3{\pm}15.9$ vs. $3.2{\pm}2.9$, respectively; p>0.001). Among groups A, B, and C, 4.9%, 31.7%, and 51.4% of the cycles, respectively, had no oocytes, and the overall rates of cycle cancellation (OPU cancellation, no oocyte, or no embryos transferrable) were 15.4%, 65.9%, and 74.3%, respectively. The fertilization rate (%) was significantly higher in group B than in group C ($73.2{\pm}38.9$ vs. $47.8{\pm}42.9$, p=0.024). The clinical pregnancy rate was significantly higher in group C than in groups A and B (44.4% vs. 27.3% vs. 9.1%, respectively, p=0.021). However, the miscarriage rate was also higher in group C than in group B (22% vs. 0%, respectively, p=0.026). Conclusion: Earlier OPU may not be effective in reducing the risk of cycle cancellation in patients with premature LH surge on hCG day. A larger scale study will be required to reveal the effectiveness of earlier ovum retrieval with premature LH surge.

• Clinical and Experimental Reproductive Medicine
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• 제30권3호
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• pp.207-215
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• 2003

Objective: To compare the pregnancy outcomes after in vitro fertilization with intracytoplasmic sperm injection (IVF-ICSI) between obstrucvtive and non-obstrucvtive azoospermia. Methods: From January 1994 to December 2002, 524 patients with obstructive azoospermia (886 cycles) and 163 patients with non-obstructive azoospermia (277 cycles) were included in this study. Microsurgical epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE) in obstructive azoospermia and TESE in non-obstructive azoospermia were perfomed to retrieve sperm, which was used for ICSI and then fertilized embryos were transferred. The results of ICSI - fertlization rate (FR), clinical pregnancy rate (CPR), clinical abortion rate (CAR) and delivery rate (DR) - were statistically analysed in obstructive versus non-obstructive azoospermia. Results: There were no differences in the number of retrieved oocytes, injected oocytes for ICSI and oocyte maturation rate. FR was significantly higher in obstructive than non-obstructive azoospermia (71.7% vs. 61.1%, p<0.001). There was no difference in CPR per embryo transfer cycle. After pregnancy was established, however, CAR was significantly higher in non-obstructive than obstructive azoospermia (25.6% vs. 12.5%, p=0.004). DR per clinical pregnancy cycle was significantly higher in obstructive than non-obstructive azoospermia (78.0% vs. 64.4%, p=0.012). In the karyotype ananlysis of abortus, abnormal karyotypes were found in 75.0% (6/8) of obstructive and 55.6% (5/9) of non-obstructive azoospermia. Conclusion: Our data show significantly higher FR in obstructive than non-obstructive azoospermia. Though there was no differrence in CPR, CAR was significantly higher in non-obstructive than obstructive azoospermia. The abortion may be related to the abnormal karyotype of embryo, but further investigations are necessary to elucidate the cause of clinical abortion in azoospermia.

• 한국수정란이식학회지
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• 제19권3호
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• pp.219-227
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• 2004

본 연구는 체외 성숙액인 TCM-199 배지를 기초로 무혈청첨가물인 PVA, PVP 및 pFF의 한정배지를 이용하여 돼지 미성숙난자의 체외성숙, 수정 및 배양 후 난자의 수정율과 배발달에 미치는 영향과 나아가 BSA 대체물로서의 이용 가능성을 알아보았다. 1. 무혈청 첨가물을 이용하여 난포란의 체외 성숙을 유기한 결과 PVA, PVP, pFF, BSA의 전체 분할율은 각각 82.4%, 78.6%, 89.4%, 90.0%로 나타났으며, GV, MI-MII율은 PVA 첨가시 각각 15.1%, 84.9%, PVP는 각각 26.5%, 73.5%, pFF는 각각 11.8%, 88.2%, BSA는 각각 11.1%, 88.9%로 PVA 혹은 pFF 첨가시에는 모두 BSA와 유사한 결과를 보였으나 PVP와는 유의적인 차이를 나타내었다 (P<0.05). 2. 체외 성숙된 난자를 수정시킨 후의 배발달율을 확인한 결과 전체 난할율은 PVA가 73%, PVP는 64.1% , pFF가 77.2%, BSA가 73%로 PVA와 pFF는 BSA와 유사하게 나타났으나, PVP는 그 발달율이 다른 처리군 들과 비교하여 유의적(P<0.05)으로 낮은 결과를 보였다. 3. 각 처리구간의 morulae와 blastocyst의 합을 비교한 결과 체외성숙 첨가물 중 PVA와 pFF는 각각 63%, 69%로 BSA(65%)와 유사한 결과를 나타내었으나 PVP는 54%로 유의적으로 낮은 결과를 나타내었다(P<0.05). 4. 전체 성숙율과 수정율을 비교해 본 결과 성숙 율은 PVA, pFF, BSA가 82.4%, 89.4%, 90.0%인 반면 PVP는 72.4%로 유의적으로(P<0.05) 낮은 결과를 보였고, 수정율은 pFF, BSA가 각각 87.1%, 89.1%로 PVA, PVP의 78.0%, 70.6%에 비해 각각 유의적(P<0.05)으로 높았다. 이상의 연구결과 돼지 난자의 체외성숙 및 배양 배지에 있어 PVA, pFF 첨가는 BSA 대체물로서 이용이 가능하나 PVP는 그 이용에 제한이 따를 것으로 판단된다.

• Clinical and Experimental Reproductive Medicine
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• 제19권1호
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• pp.41-48
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• 1992

Serum level of ${\beta}$ subunit of human chorionic gonadotropin (${\beta}-hCG$) was studied to evaluate its predictability of pregnancy outcome in 98 in vitro fertilization and embryo transfer(IVF-ET) patients using gonadotropin-releasing hormone(GnRH) agonist. Serial serum ${\beta}-hCG$ levels were established for 42 singleton pregnancies, 20 normal multiple pregnancies, 18 preclinical abortions, 14 clinical abortions and 4 ectopic pregnancies. In comparison to normal singleton pregnancies, multiple pregnancies showed significantly higher ${\beta}-hCG$ levels on the post-ET day 10 to 13 and day 24 to 25. Clinical abortions did not show significantly lower ${\beta}-hCG$ levels in early pregnancy except the post-ET day 16-17, but showed significantly lower ${\beta}-hCG$ levels from the post-ET day 22, compared with singleton pregnancies. Preclinical abortions showed significantly lower ${\beta}-hCG$ levels than those of singleton pregnancies. Ectopic pregnancies showed lower ${\beta}-hCG$ levels than those of singleton pregnancies without statistical significance. In conclusion, determination of serum ${\beta}-hCG$ level in early pregnancy is a useful tool for the prediction of preclinical abortions and multiple pregnancies and serial measurement of serum ${\beta}-hCG$ levels will be helpful in predicting clinical abortion.

• Clinical and Experimental Reproductive Medicine
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• 제29권1호
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• pp.1-12
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• 2002

Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.

• Clinical and Experimental Reproductive Medicine
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• 제36권4호
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• pp.275-281
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• 2009

목 적: 본 연구는 반복적으로 체외수정시술/난자세포질내 정자주입술을 실패했던 환자에서 배아 이식시 옥시토신 길항제의 투여가 임신율 및 착상율에 미치는 영향을 알아보고자 하였다. 연구방법: 2회 이상의 체외수정시술/난자세포질내 정자주입술을 실패했던 40명의 환자들을 대상으로 전향적 무작위 연구를 진행하였다. 과배란유도 방법으로 생식샘자극호르몬분비호르몬 길항제 다회투여법이 사용되었다. 실험군에서는 옥시토신 길항제로 atosiban (vasopressin $V_{1A}$/oxytocin antagonist)을 배아 이식 한 시간 전에 atosiban 6.25 mg을 일회 정주한 뒤, 18 mg/hour의 속도로 지속적 정맥 주입하였다. 배아 이식이 끝난 뒤 atosiban을 6 mg/hour로 감속하여 2시간 동안 추가로 정맥 주입하였다. 실험군과 대조군간의 체외수정시술 결과를 비교 분석하였다. 결 과: 실험군과 대조군간의 평균 나이, 불임 기간 및 체질량 지수와 기저 혈중 난포자극호르몬 및 에스트라디올 농도, 기저 난포강 난포의 수에 통계적으로 유의한 차이는 없었다. 또한 투여된 재조합 인간 난포자극호르몬 (rhFSH)의 총 용량과 투여 기간, 발달된 난포의 개수 및 자궁내막의 두께 역시 통계학적 유의한 차이를 보이지 않았다. 수집된 난자 및 성숙 난자의 수와 수정된 난자와 1등급 혹은 2등급의 배아 및 이식된 배아의 수에도 두 군간의 통계학적으로 유의한 차이를 보이지 않았다. 착상율을 비교하였을 때 실험군은 16.9% (11/65), 대조군은 6.0% (4/67)로 나타났고, p=0.047로 두군 간에 통계학적으로 유의한 차이를 보였다. 임상적 임신율의 경우 실험군은 40.0%로 대조군의 20.0%에 비해 높게 나타나으나 통계학적 유의성엔 도달하지 못하였다. 또한 자궁외 임신 및 유산율에 있어서는 두 군 간에 유의한 차이를 보이지 않았다. 결 론: 체외수정시술/난자세포질내 정자주입술을 시행 받는 환자들에서 배아 이식 동안 옥시토신 길항제를 투여하는 것은 자궁의 수축을 감소시켜 착상율을 증가시킬 수 있을 것으로 생각된다.

• 한국가축번식학회지
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• 제26권3호
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• pp.215-221
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• 2002

본 연구는 일정량의 항산화제(NAC, ebselen 및 GSH)와 성장인자(EGF, PDGF)의 혼합첨가가 돼지 체외수정란의 체외배양에 미치는 영향을 검토하였다. 1. NCSU 23 배양액에 NAC 1nm과 NAC에 EGF 100ng/$m\ell$비 및 PDGF 5ng/$m\ell$를 각각 혼합 첨가하여 체외배양을 실시한 결과, 상실배기 이상 발육된 체외발육율은 각각 28.1%, 32.3% 및 35.3%로서 NAC와 PDGF 혼합처리구가 대조 구보다 다소 높은 체외발육율을 나타냈으나 통계적 유의성은 없었다(P>0.05). 2. NCSU 23 배양액에 ebselen 10$\mu\textrm{m}$과 ebselen에 EGF 100ng/$m\ell$ 및 PDGF 5ng/$m\ell$를 각각 혼합첨가하여 체외발육율을 조사한 결과, 상실 배기 이상 발육된 수정란의 체외발육율은 각각 17.8%, 36.9% 및 40.3%로 ebselen과 성장인자의 혼합처리구가 대조구보다 통계적 유의하게 높은 체외발육율을 나타냈다(P<0.05). 3. NCSU 23 배양액에 GSH 100$\mu\textrm{m}$과 GSH에 EGF 100ng/$m\ell$ 및 PDGF 5ng/$m\ell$를 각각 혼합 첨가배양하여 상실배기 이상 발육된 수정란의 체외발육율은 각각 24.4%, 30.5% 및 27.7%로 GSH과 EGF 혼합처리구가 여타구보다 다소 높은 체외발육율을 나타냈지만 통계적 유의성은 없었다(P>0.05). 4. 모든 처리구에서 배반포기 수정란의 세포수는 커다란 차이가 인정되지 않았으나(P>0.05), 체외배양액에 ebselen과 성장인자를 혼합첨가하여 체외배양한 처리구에서는 대조구에 비해 통계적으로 유의하게 높은 세포수를 나타냈다.(P<0.05).

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.122-129
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• 2020

Objective: The survival of a semi-allogeneic fetus depends on several immunological mechanisms, and it has been suggested that recurrent pregnancy loss (RPL) could develop as a result of one or more immunological abnormalities. Methods: Compatibility between partners for human leukocyte antigen (HLA) genotypes and the relationships between maternal killer-cell immunoglobulin-like receptor (KIR) and paternal HLA-Bw4/Bw6 and HLA-C1/C2 supra-groups were investigated in 25 couples with RPL in comparison to healthy couples with children. HLA and KIR genotyping was performed using polymerase chain reaction with sequence-specific primers and/or sequence-specific oligonucleotides. Results: HLA class I incompatibility between partners, especially in HLA-B alleles, was more common in the RPL group (p= 0.01). HLA-C2 homozygosity was more frequent in the male partners of RPL couples than in other groups (p= 0.03). The KIR2DL5 gene frequency was significantly higher in both the female and male partners of RPL couples, whereas the KIR2DS3 gene frequency in male partners of RPL couples was significantly reduced (p= 0.03). The presence of KIR2DL3 in women with RPL was correlated with the presence of HLA-C2 alleles in their spouses (p= 0.03). Conclusion: Our data from a Turkish population suggest that male HLA-C2 homozygosity may play an important role in RPL. Additionally, an incidental match between male HLA-C2 and female HLA-C1 ligand KIR receptors might perturb the balance between activatory and inhibitory KIR-ligand interactions during pregnancy in couples affected by RPL. The roles of orphan KIR2DL5 and orphan KIR2DS3 in RPL remain obscure.