• 한국발생생물학회지:발생과생식
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• 제4권2호
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• pp.203-213
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• 2000

본 연구에서는 반응성 산소족이 수정능력획득, 칩체반응에 미치는 영향을 알아보고자 반응성 산소족으로 superoxide anion은 xanthine (X) -xanthine oxidase (XO) system을, hydroperoxide는 $H_2O$$_2$를 농도별로 처 리하였으며, nitric oxide (NO)는 NO donor인 sodium nitroprusside (SNP)를 처리하였다 또한 남성불임요인의 하나로 알려진 leukocytospermia에 대한 영향을 알아보기 위해 lymphocyte를 농도별로 처리하였고, 일반적인 배양기내 산소농도인 20% $O_2$농도를 생체내 농도와 유사한 5% $O_2$ 농도로 낮추었을 때 의 결과를 알아보고자 하였다. 수정능력 획득 정도와 첨체반응률을 알아보기 위해 chlortetracycline (CTC) 염색방법을 이용하였다. 지질과산화 정도는 정자내 malondialdehyde (MDA)의 생성량을 흡광기를 이용하여 정량하였다. $H_2O$$_2$, X-XO, SNP와 lymphocyte 처리군은 1시간 배양시에 수정능력획득률이 유의하게 증가하였으나, 저산소처리군에서는 차이가 없었다. 저 농도의 $H_2O$$_2$를 처리한 경우에는 지질과산화 정도가 감소하였으나, 고 농도에서는 대조군에 비해 유의하게 증가하였다. 고 농도의 Iymphocyte를 처리한 경우에는 1시간 처리시에 지질과산화가 유의하게 증가하였으나, 처리된 산소농도에 따른 지질과산화의 차이는 없었다. 첨체반응률의 경우, 처리한 모든 반응성 산소족에서 대조군에 비해 높은 첨체반응률을 확인하였다. X(100 $\mu$M)-XO(100mIU)의 경우가 가장 높은 첨체반응률을 나타내었다. 이러한 결과들은 반응성 산소족이 수정능력획득, 지질과산화 그리고 첨체반응에 영향을 미치는 것을 확인하여 주었다. 또한 반응성 산소족이 생성된 경우에 수정능력획득이 보다 빠르게 진행되어지는 것은 반응성 산소족이 정자의 과운동성과 수정능력획득의 중요한 조절자임을 시사한다고 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.23-29
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• 2000

Objective: Zona pellucida (ZP) has been thought to be the barrier of egg to sperm penetration before and after fertilization. The phenomenon of ZP hardening has been considered as a post-fertilization event until now, and it is generally accepted that it is caused by the secretory products of cortical granules released during the cortical reaction. Hardening of ZP could occur "spontaneously" in mammalian oocytes in standard culture conditions, and that it is probably not a consequence of cortical reaction. The purpose of our study was to investigate the effect of human amniotic fluid (HAF) on nuclear maturation (NM) and fertilization ability of mouse immature oocytes. Methods: HAF was obtained from patients undergoing amniocentesis at $16{\sim}20$ weeks of gestation. HAF from five to ten patients was centrifuged and the supernatants was pooled. Cumulusenclosed mouse immature oocytes were incubated in the medium containing HAF, and examined to confirm NM and fertilization. Female ICR mice (about 3 weeks old) were stimulated with 7.5 IU PMSG. Immature oocytes were isolated at $48{\sim}52$ hrs post PMSG injection and cultured in TCM-199 supplemented with 20% HAF for 18 hrs. FBS was used as a control for the examination. Matured oocytes (MII) were fertilized with sperms collected from the epididymis of male mice (over 10 weeks old). Fertilization was in conducted T6 medium containing 15 mg/ml BSA, and confirmed at 6 hrs post-insemination. Fertilization rate was assessed in zona-intact or zona-free oocytes (denuded by trypsin). Evaluation of NM and fertilization was carried out by rapid staining method. ZP hardening was evaluated by incubating cumulus cell-free mature oocytes in 0.001% chymotrypsin at $37^{\circ}C$ for 10 min. Results: There was no significant difference between the effects of HAF (86.6%) and FBS (87.7%) supplements on NM of immature oocytes. When maturation medium was supplemented with HAF, total fertilization rates (7%) were significantly lower (p<0.01) than that of FBS (85.1%). In HAF group, fertilization rate was increased (p<0.01) in zona-free oocytes (7% versus 100%). The resistance of mouse oocyte ZP to digestion by chymotrypsin after maturation in vitro was significantly higher (p<0.01) in HAF group (86.7%) than in FBS (6.7%). To culture oocytes in FBS were very effective in preventing ZP hardening. However cultured oocytes in HAF showed high rate of ZP hardening (p<0.01). Conclusions: These results suggest that HAF can be used as a supplement for the NM of mouse immature oocytes in vitro. However, HAF induces spontaneous hardening of ZP of mouse immaure oocytes during maturation in vitro.

• Clinical and Experimental Reproductive Medicine
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• 제38권1호
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• pp.31-36
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• 2011

Objective: To determine whether the serum ${\beta}$-human chorionic gonadotropin (hCG) profile following preimplantation genetic diagnosis (PGD) is lower than that of intracytoplasmic sperm injection (ICSI) cycles. Methods: A total of 129 PGD cycles and 1,161 age-matched ICSI cycles, which resulted in pregnancy (serum ${\beta}-hCG{\geq}5$ mIU/mL) on post-ovulation day (POD) 12 were included. We compared the mean serum ${\beta}$-hCG levels on POD 12, 14, 21, and 28, doubling time of serum hCG, and created a cut-off value for predicting a singleton pregnancy in each group. Results: The mean serum ${\beta}$-hCG concentration of the PGD group was significantly lower than that of the control group on POD 12, 14, and 21. The doubling time of serum ${\beta}$-hCG at each time interval showed no significant difference. The cut-off-value of serum ${\beta}$-hCG for predicting a single viable pregnancy was 32.5 mIU/mL on POD 12 and 113.5 mIU/mL on POD 14 for the PGD group, which was lower than that for the control group. Conclusion: Blastomere biopsy may decrease the ${\beta}$-hCG-producing activity of the trophoblasts, especially in early pregnancy. Setting a lower cut-off value of serum ${\beta}$-hCG for predicting pregnancy outcomes in PGD may be needed.

• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.281-292
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• 1997

The objective of this study was to compare retrospectively the survival and pregnancy rates(PR) of cryopresered-thawed embryos obtained from intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (IVF). Ninety-six cycles of cryopresered-thawed embryo transfer (ET) were performed in 79 patients from June, 1996 to September, 1997 and grouped as followings: 20 cycles (16 patients) inseminated by ICSI (ICSI Group) and 76 cycles (63 patients) by conventional IVF (IVF Group). Slow-freezing and rapid-thawing protocol was used with 1.5M propanediol (PROH) and 0.1M sucrose as cryoprotectant. All embryos were frozen-thawed at the two pronuclear (2 PN) stage excluding four cycles in which the early cleavage stage embryos were frozen, and allowed to cleave in vitro for one day before ET. The duration from freezing to thawing was comparable in both groups ($mean{\pm}SD$, $112.1{\pm}80.0$ vs. $124.8{\pm}140.1$ days). The age of female ($31.2{\pm}3.4$ vs. $32.6{\pm}3.3$ years) and the endometrial thickness prior to progesterone injection ($9.4{\pm}2.0$ vs. $9.3{\pm}1.8$ mm) were also comparable in both groups. There was no significant difference in the outcomes of cryopreserved-thawed ET between two groups: survival rate ($85.2{\pm}16.1%$ vs. $82.2{\pm}19.7%$), cleavage rate ($96.9{\pm}6.7%$ vs. $94.7{\pm}13.0%$), cumulative embryo score (CES, $54.5{\pm}31.1$ vs. $49.0{\pm}20.0$), preclinical loss rate (5.0% vs. 5.3%), clinical miscarriage rate (0% vs 29.4%), clinical PR per transfer (35.0% vs. 22.4%), implantation rate (9.9% vs. 5.6%), and multifetal PR (42.9% vs. 17.6%). In conclusion, human embryos resulting from ICSI can be cryopreserved-thawed and transferred successfully, and the survival rate and PR are comparable to conventional IVF.

• 한국어류학회지
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• 제21권4호
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• pp.247-252
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• 2009

멸종위기 담수어류인 얼룩새코미꾸리 Koreocobitis naktongensis의 난 발생 과정 및 자치어 형태 발달을 조사하였다. 인간 융모성 성선자극호르몬(HCG)을 10 IU/g의 농도로 암컷의 복강에 주사하여 성숙란을 얻었으며, 건식법으로 정자와 수정시켰다. 수정란의 난경은 1.0~1.2 mm로 유구는 존재하지 않았다. 배체는 수온 $20{\pm}1^{\circ}C$에서 수정 38시간 후부터 부화하기 시작하였다. 부화 직후 자어의 전장은 2.5~2.8 mm로 입과 항문은 아직 열리지 않았다. 부화 후 7일째 자어는 전장 4.5~5.2 mm로 난황은 완전히 흡수되었다. 부화 후 50일째 모든 지느러미가 완성되는 치어기로 이행하였고, 이때의 전장은 21~27 mm였다.

• Clinical and Experimental Reproductive Medicine
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• 제25권3호
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• pp.245-249
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• 1998

White blood cells (WBCs) are present in most human ejaculates, but abnormally high concentration of seminal leukocytes may reflect an underlying pathological condition. The World Health Organization (WHO) has defined leukocytospermia as status of more than $10^6$ WBC/mL of semen. The purpose of this study was firstly, to compare the outcomes between conventional IVF and ICSI in leukocytospermia, and secondly, to investigate whether ICSI may be an alternative treatment for patients with leukocytospermia. Total 121 cycles of conventional IVF and ICSI candidates underwent IVF cycles at PL Infertility Clinic. Semen Parameters including concentration, motility, morphology of spermatozoa and concentration of leukocytes were assessed from the raw ejaculates. There was no difference in sperm concentration, motility and morphology. The rates of fertilization and good embryo development from ICSI were significantly higher than those from conventional IVF in leukocytospermia (63.9% & 48.6%, respectively for ICSI group and 33.4% & 24.1%, respectively for IVF group, p<0.001). The pregnancy rate after ICSI was also higher than that from conventional IVF (34.3% vs 21.6%, p<0.05). These results indicate that the presence of seminal leukocytes ($>1\times10^6$ WBC/mL of semen) is adversely related with fertilization, embryo development and pregnancy rate. Therefore the measurement of seminal leukocytes in routine semen analysis appears to be of prognostic value with regard to male fertilizing potential. In conclusion, it is suggested that ICSI is an alternative choice of treatment for patients with leukocytospermia.

• Clinical and Experimental Reproductive Medicine
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• 제25권3호
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• pp.269-275
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• 1998

It has been reported that the $Ca^{2+}$-ATPase and the $Ca^{2+}-Na^+$ exchanger play an important role for the regulation of intracellular $Ca^{2+}$ in somatic cells, the $Ca^{2+}$-ATPase located in the plasma membrane helps the $Ca^{2+}$ concentration in maintain low $[Ca^{2+}]_i$. Roldan & Fleming reported that the spermatozoan $Ca^{2+}$-ATPase plays an important role in the capacitation and acrosome reaction. We used to assess $Ca^{2+}$ changes by chlortetracycline (CTC) patterns in the capacitation and acrosome reaction of human and hamster spermatozoa. In the present study applying quercetin which has been known as an ATPase antagonist, the enzymatic effect of $Ca^{2+}$-ATPase on capacitation and acrosome reaction was found to be remarkable: a significant increase of the transformation from the original type to the B type and the AR type of spermatozoa. This finding suggests that $Ca^{2+}$-ATPase play an important role in the efflux and the influx of the $Ca^{2+}$ which have been known to be an essential factor for the capacitation and acrosome reaction, and that the inhibitory action of the $Ca^{2+}$-ATPase might be a prerequsit step toward the capacitation and acrosome reaction. In conclusion, this study suggest the considerable evidence as follows: the increment of the intracellular $Ca^{2+}$ concentration occurred by controlling the slope of $Ca^{2+}$ concentration through $Ca^{2+}$-ATPase activites in both the intracellular and extracellulr fluid may be important procedures for the capacitation and the acrosome reaction, and finally for fertilization of the sperm and ovum.

• Clinical and Experimental Reproductive Medicine
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• 제3권2호
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• pp.35-42
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• 1976

The human semen ejaculated in a form of liquid state, coagulates immediately after ejaculation, and then liquefies again. However, the mechanisms of neither coagulation and liquefaction of semen have not been explained clearly so far, and very limited numbers of report are available, although the spermatology and andrology made rapid progress. This clinical study has been undertaken to investigate the liquefaction phenomena and practicability of the results might be applied to fertility and infertility problems. As a preliminary study, in this report the liquefaction time of various semen groups is measured and analysed. The following results are obtained: 1. An average liquefaction time of semen of a total of 60 subjects: 25 minutes. 2. An average liquefaction time of semen according to sperm count: 1) Normospermia group (20 cases): 34 minutes. 2) Oligospermia group (20 cases): 21 minutes. 3) Azoospermia group (20 cases): 20 minutes. 3. An average liquefaction time of semen according to abstinence period: 1) Less than 3 days group (30 cases): 22 minutes. 2) More then 5 days group (30 cases): 28 minutes. In conclusion: 1. The liquefaction time of semen of the normospermia group is longer than oligospermia group or azoosermia group. 2. The liquefaction time of semen may not be greatly influenced by the various factors such as abstinence period, semen volume, semen pH, age of the subjects and so on. 3. In routine semen analyses, it is recommended to begin the analysis at least 25 minutes after the ejaculation. 4. Further studies are required in conjunction with practical application of liquefaction mechanism in infertility and fertility control.

• Clinical and Experimental Reproductive Medicine
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• 제13권2호
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• pp.121-127
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• 1986

We have reviewed 59 cases of patients amoung 65 cases who underwent IVF and ET with reasonable indications irom 1984 and the results as follows. 1. Major indications for IVF and ET were tubal factor (40.7%), unexplained infertility (25.4%), endometriosis (15.3%), failed AID and AIH (10.1 %), and sperm abnormality (8.5%). 2. For superovulation of human oocytes, l00mg of clomiphene citrate and 75 IU of HMG used. The monitoring of oocyte maturation was bone by ultrasound examination and serum 17-${\beta}$ estradiol, LH values. The peak $E_2$ value was 956.36${\pm}$702.13 pg/ml. 3. The oocytes were obtained by laparoscopy 24-36 hours after the injection of HCG. 4. The mean numbers of follicles at laparoscopy was 3.06 and the successful rate of laparoscopy was 79.7%. 5. And 165 follicles were aspirated from which 98 oocytes were recovered, 59.4% of all follicles had at least one oocyte aspirated. 21.4% of the eggs were mature, 52.0% were moderate, 26.5%. were immature. 6. 67.3% of oocytes were cleaved and were transferred at 4-6 cell stages. 7. Four pregnancies including one chemical pregnancy and one spontaneous abortion were established by ${\beta}$-subunit, u-hCG and ultrasound examinations.

• Clinical and Experimental Reproductive Medicine
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• 제44권3호
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• pp.141-145
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• 2017

Objective: Delaying embryo transfer (ET) enables us to select among the embryos available for transfer and is associated with positive effects on implantation and pregnancy outcomes. However, the optimal day for ET of human cleavage-stage embryos remains controversial. Methods: A retrospective study of 3,124 in vitro fertilization/intracytoplasmic sperm injection cycles (2,440 patients) was conducted. We compared the effects of day 2 and 3 ET on rates of implantation and pregnancy outcomes between young maternal age (YMA; < 38 years old, n = 2,295) and old maternal age (OMA; ${\geq}38years\;old$, n = 829) patient groups. Results: The YMA and OMA groups did not differ in terms of patient characteristics except for the proportion of unexplained factor infertility, which was significantly greater in the OMA group, and the proportion of arrested embryos, which was significantly greater in the YMA group. However, the biochemical pregnancy, clinical pregnancy, ongoing pregnancy, abortion, and implantation rates per cycle were not significantly different between day 2 and 3 ET in the YMA group or the OMA group. Conclusion: We suggest that offering patients the opportunity to decide which day would be suitable for ET could be part of a patient-friendly protocol that takes into consideration an infertile woman's circumstances and work schedule by allowing ET to be performed on day 2 instead of the traditional transfer on day 3.