• Clinical and Experimental Reproductive Medicine
• /
• 제37권1호
• /
• pp.49-56
• /
• 2010

목 적: 저성선자극호르몬 성선저하증 환자에서 보조생식술의 임신 결과에 대해 알아보고자 하였다. 연구방법: 저성선자극호르몬 성선저하증으로 진단받고 본원에서 보조생식술을 시행받은 23명을 연구군으로, 동일기간 난관요인으로 보조생식술을 시행받은 이들 중 연구군과 연령 및 체질량지수가 일치하는 120명의 여성을 대조군으로 설정하여, 이들의 의무기록을 후향적으로 열람하였다. 보조생식술 관련 여러 계측치 및 임신율, 유산율, 출산율 등을 비교 분석하였다. 결 과: 연구군의 평균 연령은 $32.7{\pm}3.3$세였고, 평균 체질량지수는 $21.0{\pm}3.2kg/m^2$였다. 생리주기 제 2~3일에 측정한 황체형성호르몬과 난포자극호르몬은 각각 $0.61{\pm}0.35$, $2.60{\pm}2.35$ mIU/ml였고, 에스트라디올은 $10.13{\pm}8.17$ pg/ml이었다. 난소자극 주기에서 사용된 생식샘자극호르몬의 총 양과 투여기간 및 hCG 투여일의 $E_2$ 수치는 연구군에서 유의하게 높았다. 보조생식술 방법에 따라 분석한 결과, 체외수정 및 배아이식 (IVF-ET) 주기에서는 연구군에서 자궁내막두께와 수정율, 출산율이 유의하게 낮았고, 유산율은 유의하게 높았으며, 그 외 난소자극 및 인공수정 (SO-IUI) 및 동결보존배아이식 (FET) 주기에서는 두 군간에 유의한 차이를 보이지 않았다. 결 론: 저성선자극호르몬 성선저하증 여성에서 생식샘자극호르몬 치료를 통한 전반적인 보조생식술의 임신율은 22.0%로 대조군의 그것과 유사하지만, 이를 위해서는 더 많은 용량의 호르몬이 필요하다. 연구군의 IVF-ET의 경우, 주기 중 현저하게 높은 $E_2$ 수치와 유의하게 얇은 자궁내막을 보이며, 더 높은 유산율과 더 낮은 생존출산율을 보여, 이의 극복을 위해 자궁내막 수용성 개선방안에 대한 연구가 추가로 필요하다.

• Clinical and Experimental Reproductive Medicine
• /
• 제23권3호
• /
• pp.319-326
• /
• 1996

본 연구는 인간의 체외수정 program으로 부터 생산된 여분의 배반포기 배를 이용하여 현미경에 의한 형태학적 판정과 differential labelling 기법을 이용한 세포 수의 상관관계를 조사하고자 실시하였다. 공시된 인간 배반포기 배는 체외수정 후 5일째에 36명의 환자로부터 76개를 얻어 배반포강의 확대와 투명대 두께의 감소를 기준으로 early (ErB), early expanding (BEB), middle expanding (MEB) 및 expanded blastocyst (EdB)로 구분하였다. 분류된 배반포기 배의 크기와 투명대의 두께를 micrometer로 측정하였을 때, 그 크기는 각각 $148.8-217.6{\mu}m$, $1.2-14.4{\mu}m$로 나타나 같은 배양조건에서 생산된 배아라도 그 차이는 크게 나타났다. Hoechst 염색을 이용하여 배반포기 배의 총 세포수를 조사하였을 때, 체외수정 후 5일째 생산된 배반포기 배는 ErB $(39.1{\pm}3.6)$ 에서 EdB $(89.6{\pm}3.3)$ 로 진행되는 동안 두배에서 세배정도 증가되는 양상을 나타내었다. 또한, differential labelling기법 을 이용한 배반포기 배의 inner cell mass (ICM) 와 trophectoderm (TE) 세포수 조사 결과, 각각 $11.9{\pm}1.8-22.2{\pm}4.3$, $24.5{\pm}3.6-70.0{\pm}7.7$을 나타내어 발달이 진행될수록 ICM과 TE세포수도 증가되는 것을 알 수 있었다. 특히, 형태학적으로 약한 ICM으로 판정된 EdB의 경우 differential labelling 후, 역시 적은 ICM 세포수를 나타내어 형태학적 판정과 세포수간에 밀접한 상관관계가 있음을 알 수 있었다. 따라서, ICM과 TE를 differential labelling하는 기법은 인간 배반포기 배의 quality를 평가하는데 매우 유용한 기법으로 형태학적인 구분과 병용된다면 인간 배반포기 배 이식 program의 임신율 증진을 위한 배아 선별의 중요한 자료로서 이용될 수 있다는 것을 시사한다.

• Clinical and Experimental Reproductive Medicine
• /
• 제25권3호
• /
• pp.251-260
• /
• 1998

It is well known that the clinical test for responsibility of accurate fertilization capacity in male partners is very important to diagnose and treat the infertility. However, it has been reported that the traditional semen analysis cannot accurately predict fertilization and pregnancy potential. The present study was performed to evaluate the acrosomal reaction to ionophore challenge (ARIC) test as a prognostic indicator for fertilization of sperm and oocyte in an in vitro fertilization and embryo transfer (IVF-ET) program. From March 1996 to Februry 1997, 30 couples undergoing IVF program were allocated to this study group. All female partners in the study group were 35 years old or less and their serum level of basal follicle stimulating hormone (FSH) and estradiol $(E_2)$ were normal. All the male partners have normal parameters of semen analysis. The ARIC tests were performed on the day of ovum pick up and in vitro insemination in all the male partners. The controlled ovarian hyperstimulation (COH) using luteal long protocol of gonadotropin releasing hormone (GnRH) agonist was used in all couples for IVF-ET. The acrosomal reaction with $10{\mu}l$ of 10% DMSO was induced spontaneously in $10.1{\pm}9.8%$, and acrosomal reaction with calcium ionophore A 23187 was induced in $27.4{\pm}18.1%$, and the ARIC value was $17.4{\pm}16.2%$. There were no significant correlation between the ARIC value and the fertilization rate ($r^2$=0.044, p=0.268). There were also no significant correlation between the ARIC value and the percentage of the grade I, II embryos ($r^2$=0.046, p=0.261). On the basis of above results, it was suggested that ARIC test might not be a useful prognostic indicator for fertilization in IVF-ET in male partners with normal parameters of conventional semen analysis. We guessed that IVF-ET could be performed to the patients primarily without universal appilcation of ARIC test to all male partenrs, and if fertilization failure occurs, the micro assisted fertilization (MAF) such as intracytoplsmic sperm injection (ICSI) might be used as an alternative mode of treatment with acceptable success rate.

• Clinical and Experimental Reproductive Medicine
• /
• 제29권1호
• /
• pp.45-56
• /
• 2002

Objectives: Recently, recombinant FSH (rFSH) has been manufactured using a Chinese hamster ovary cell line transfected with the gene encoding human FSH. Both rFSH and urinary gonadotropin (uFSH) could be used for controlled ovarian hyperstimulation (COH). However, uFSH implies a number of disadvantages, such as batch-to-batch inconsistency, no absolute source control, dependence on large amounts of urine, low specific activity, and low purity. The purpose of this study was to evaluate the efficacy of rFSH in human IVF-ET program. Materials and Methods: A total of 508 infertile women was enrolled in this study. They are classified into rFSH group (n=177) or uFSH group (n=331), and all of them were matched by age and cause of infertility in same period. The $Puregon^{(R)}$ (Organon, Holland) was used as rFSH, and the Metrodin-$HP^{(R)}$ (Serono, Switzeland) and $Humegon^{(R)}$ (Organon, Holland) was used as uFSH. We subdivided the patients into three age groups. The outcomes of IVF-ET program were analyzed using the statistical package for social sciences (SPSS). Results: There was no significant differences in the level of estradiol on hCG injection day, the numbers of retrieved oocytes, matured oocytes, fertilized oocytes, transferred embryos, frozen embryos between the two groups. The total dose (IU) of gonadotropin for COH was significantly lower in the rFSH group compared to uFSH group ($1339{\pm}5491.1$ vs $2527.8{\pm}1075.2$ IU, p<0.001). Clinical pregnancy rate per embryo transfer in the rFSH group showed increasing tendency, compared to the uFSH group, but there was no statistical significance (35.2% vs 29.3%). Our results demonstrated that the relative efficiency of rFSH compared with uFSH is higher in older patients. Conclusions: The ovarian stimulatory effect and clinical outcome of recombinant FSH was similar to that of the urinary gonadotropin. The IVF-ET cycles with significantly lower dose of gonadotropin in rFSH group showed comparable results. Therefore, we suggest that recombinant FSH is more potent and effective than urinary gonadotropin.

• 식물조직배양학회지
• /
• 제26권3호
• /
• pp.189-195
• /
• 1999

녹색꽃양배추의 약배양에 영향을 미치는 제 조건을 규명하고 다양한 유전적인 특성을 가진 품종들을 대상으로 다수의 계통을 유기하여 신품종 육성 소재로 이용하고자 실험한 결과는 다음과 같다. 배지시험 결과 B5배지에 NAA 0.1 mg/L + 2.4-D 0.1mg/L + 당 10% 처리구에서 배상체 발생율이 가장 높았으며 액체배지가 고체배지보다 5배 정도 배발생에 효과적이었다. 약접종 직후 35$^{\circ}C$ 고온처리 일수에 따른 시험에서는 1일처리가 가장 효과적이었으며 시일이 경과할수록 배상체 발생율이 현저히 저하되었다 품종간 배상체 유기율은 Green valient 2.8%에서 Haisi의 21%까지 차이가 심하였으나 이배체율은 62~74%로 품종간 큰 차이를 나타내지 않았다. Rokguray와 Haisi두 품종 약유래 계통의 포장특성 검정결과 초장, 엽수, 측지수, 화뢰의 지름은 다양한 변이를 보였으며 두 품종에서 유기된 계통들 중 조숙이며 초장이 크고 화뢰의 지름이 큰 계통과 중만생이며 엽수가 많고 화뢰의 지름이 작은 계통을 각각 5계통, 7계통을 선발하여 F$_1$육성 소재로 이용하였다.

• 한국가축번식학회지
• /
• 제20권2호
• /
• pp.207-214
• /
• 1996

본 연구는 미세조작된 수정란의 초급속 재동결 후 발생능을 조사하기 위해서 수행하였다. 먼저 4세포기 생쥐 수정란으로부터 할구세포 한 개를 떼어내고 이들 수정란을 동결액에 넣어 상온에서 2.5분간 평형시킨 다음, 0.25ml straws에 넣어 곧바로 액체질소에 침지시켰다. 4세포기 수정란의 동결액으로는 4.0M(ethylene glycol 및 0.25M sucrose가 함유된 dPBS를 사용하였따. 상실배기 수정란의 동결을 위해서는 항동해제로 4.0M ethylene glycol 대신에 5.0M glycerol을 사용하였다. 융해후 biopsied 4세포기 수정란을 M16 배양액에서 상실배기까지 발달시킨 다음 상실배기용 동결액을 이용하여 초급속 재동결을 실시하였다. 재동결융해 후 발달한 biopsied 배반포기 수정란을 대리모에 이식하여 이들 수정란의 생존성을 검토하였다. 제1차 동결후 biopsied 수정란의 체외발달률은 78%로서 정상적인 수정란의 발달율(91%) 보다 낮은 성적으로 보여주었으나 (P<0.01), 이들 수정란의 이식 후 임신률은 각각 25 및 30%로서 두 실험군간에 차이가 인정되지 않았다. 그리고 biopside 상실배기 수정란의 초급속 재동결후 체외 발달률 및 임신률은 각각 89와 27%로서 biopsied 되지 않은 수정란의 성적 (각각 95 및 28%)과 유사하였다. 이러한 결과를 종합하여 볼 때, 본 연구에서 사용된 초급된 재동결 과정이 미세조작된 생쥐수정란의 생존성에 영향을 미치지 않는 것으로 나타났다.

• Clinical and Experimental Reproductive Medicine
• /
• 제31권2호
• /
• pp.133-139
• /
• 2004

Objective: To investigate the association of FSH receptor (FSHR) polymorphism at position 680 with outcomes of controlled ovarian hyper-stimulation for IVF-ET in Korean women. Design: Genetic polymorphism analysis. Materials and Methods: The FSHR polymorphism was analyzed by PCR-RFLP in 172 ovulatory women below the age of 40 year. Patients with polycystic ovary syndrome, endometriosis, or previous history of ovarian surgery were excluded. Results: Genotype distribution was 41.9% for the Asn/Asn, 47.7% for the Asn/Ser, and 10.5% for the Ser/Ser FSHR genotype group. There was no difference in age of subjects and infertility diagnosis between genotype groups. When the patients were grouped according to their FSHR genotype, the basal levels of FSH (day 3) were significantly different among the three groups ($6.0{\pm}0.3\;IU/L$ (mean $\pm$ SEM), $5.8{\pm}0.3\;IU/L$, and $8.6{\pm}1.2\;IU/L$ for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively, p=0.002). The Ser/Ser group showed a higher total doses of gonadotropins required to achieve ovulation induction, and a lower serum estradiol levels at the time of hCG administration compared with other two groups, but the differences were of no statistical significance. The numbers of oocytes retrieved were significantly different among the three groups ($8.6{\pm}0.8$, $9.9{\pm}0.6$, and $6.3{\pm}0.9$, for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively, p=0.049). Clinical pregnancy rates were 42.4%, 25.9%, and 29.4% for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively. Conclusion: Homozygous Ser/Ser genotype of FSHR polymorphism at position 680 was associated with decreased ovarian response to gonadotropin stimulation for IVF-ET.

• Clinical and Experimental Reproductive Medicine
• /
• 제27권1호
• /
• pp.67-74
• /
• 2000

Objective: This study was conducted to investigate the effect of vitrification on the implantation and the pregnancy of human blastocysts. Method: The transfer of the frozen-thawed blastocysts by the slow freezing or vitrification was performed between January 1998 and July 1999. The zygotes derives from IVF were cocultured with cumulus cells in YS medium containing 20% hFF for 5days. Two or three of the best balstocysts produced on day 5 were transferred into the uterus, and then supernumerary blastocysts were randomly divided into two groups. One was frozen by slow freezing and the other was frozen by vitrification method. The slow freezing procedure was performed in two steps (5% glycerol and 9% glycerol + 0.2 M sucrose for 10 min, respectively) using programmed freezer ($-2^{\circ}C$/min to $-7^{\circ}C$, manual seeding at $-7^{\circ}C$, $-0.3^{\circ}C$/min to $-38^{\circ}C$ and plunged into $LN_{2}$). The blastocysts frozen by slow freezing were thawed at $36^{\circ}C$ then removed glycerol in 7 steps. The vitrification procedure was performed in three steps (10% glycerol for 5 min, 10% glycerol + 20% ethylene glycol for 5 min, 25% glycerol + 25% ethylene glycol and directly $LN_{2}$ within 1 min). The blastocysts frozen by vitrification were thawed at $20^{\circ}C$ water then removed cryoprotectant in 3 steps. In each group, thawed blastocysts were cocultured with cumulus cells in YS medium containing 20% hFF for 18h and transferred into the uterus. The implantation rate was evaluated per transferred blastocysts and the pregnancy rate was evaluated per transfers. Results: The survival rate of vitrified group (74.5%) was higher than slow freezing group (68.0%), but not significant. When 98 thawed blastocysts of vitrification were transferred in 40 cycles, 19 pregnancies (clinical pregnancy rate; 47.5%) were established. One miscarriage occurred in the eighth week of pregnancy (ongoing pregnancy rate; 45.0%). 7 pregnancies were ongoing, 11 pregnancies went to term, and 16 healthy infants were born. The Implantation rate was 31.6%. These results were higher than those obtained by the slow freezing (clinical pregnancy rate; 40.3%, ongoing pregnancy rate; 32.5% and implantation rate; 25.3%), but not significant. Conclusion: Vitrification is a simple, quick and economical method when compared to slow freezing. It will be chosen as a good method of human embryo freezing in IVF-ET programs.

• 한국가축번식학회지
• /
• 제10권2호
• /
• pp.192-203
• /
• 1986

Two experiments in this study were designed to compare the potential for in vitro capacitation and in vitro fertilization of ejaculated sperm among individual rabbit bucks. In experiment 1, for in vitro capacitation, the ejaculated sperm were preincubated in DM for 12 hr or 18 hr after HIS treatment, then 12 hr -or 18 hr- preincubated sperm were incubated with superovulated rabbit ova in a 5% CO2 incubator for 36 hr at 38$^{\circ}C$, and a part of cleaved ova was transferred to the recipient does for implantation of embryo. In experiment 2, effect of lysolecithin addition to preincubation medium on induction of accelerated in vitro capacitation and in vitro fertilization of individual rabbit sperm was studied. Experiment 1; 1. Percent acrosome reaction of sperm, noted after staining, after 12 hr or 18 hr preincubation ranged from 52.5 to 76.0% and from 67.5 to 90.0%, respectively and sperm motility index of these sperm ranged from 20.0 to 47.5 for 12 hr-preincubated sperm and from 15.0 to 37.5 for 18 hr- preincubated sperm. There was no a certain relation between percent acrosome reaction and sperm motility index. 2. In vitro fertilization rate (cleavage rate) of in vitro capacitated sperm varied widely among individual bucks, ranging from 0 to 47.8% for 12 hr - preincubated sperm and from 0 to 60.9% for 18 hr -prein- cubated sperm. Cleavage rate of 18 hr - preincubated sperm was higher and faster than that of 12 hr - preincubated sperm. 3. Eight of 44 in vitro fertilized embryos transferred into 6 recipients were implanted in 4 recipients (66.7%) up to day 15 and implnatation rate was 18.2%. Experiment 2; 1. The percent acrosome reaction of sperm before and after 4 hr preincubation in DM without lysolecithin varied significantly among individual bucks, ranging from 0.4 to 18.4% and from 1.7 to 37.4%, respectively and percent acrosome reaction of sperm at 30 min after addition of 60${\mu}$g/ml lysolecithin also was significantly different among bucks, ranging from 19.2 to 67.1%. 2. Effect of accelerated acrosome reaction following lysolecithin addition was more considerable in the individuals showed less percent acrosome reaction before and after 4 hr preincubation. Percentage of motile sperm and motility score showed a trendency towards a decrease with increase of preincubation time and time after lysolecithin addition. 3. In vitro fertilization rate (cleavage rate) at 24 hr postinesmination with pooled sperm were treated to 60 $\mu\textrm{g}$/ml lysolecithin for 30 min after 4 hr preincubation was 24.6%, a higher rate than 13.2% for control. While 80 $\mu\textrm{g}$/ml lysolecithin-added sperm showed a lower cleavage than control and 60$\mu\textrm{g}$/ml-added sperm at both 24 hr and 48 hr postinsemination. These results from 2 experiments suggest that more useful preincubation time for the in vitro capacitation of ejaculated rabbit sperm is 18 hr in DM after HIS treatment, although there is wide variation in vitro capacitation and in vitro fertilization rate among individual bucks, and lysolecithin addition to at least 4 hr - preincubated sperm in DM can result in almost same in vitro fertilization rate as that of 18 hr - preincubated sperm in the experiment 1.