• Clinical and Experimental Reproductive Medicine
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• 제39권4호
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• pp.166-171
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• 2012

Objective: We compared the assisted reproductive technology (ART) outcomes among infertile women with polycystic ovary syndrome (PCOS) treated with IVM, conventional IVF, GnRH agonist, and GnRH antagonist cycles. Methods: The prospective study included a total of 67 cycles in 61 infertile women with PCOS. The women with PCOS were randomized into three IVF protocols: IVM/IVF with FSH and hCG priming with immature oocyte retrieval 38 hours later (group A, 14 cycles), GnRH agonist long protocol (group B, 14 cycles), and GnRH antagonist multi-dose flexible protocol (group C, 39 cycles). IVF outcomes, such as clinical pregnancy rate (CPR), implantation rate (IR), miscarriage rate (MR), and live birth rate (LBR), were compared among the three groups. Results: Age, BMI, and basal FSH and LH levels did not differ among the three groups. The number of retrieved oocytes and 2 pronucleus embryos was significantly lower in group A compared with groups B and C. The CPR, IR, MR, and LBR per embryo transfer showed no differences among the three groups. There was no incidence of ovarian hyperstimulation syndrome in group A. Conclusion: The IR, MR, and LBR in the IVM cycles were comparable to those of the GnRH agonist and GnRH antagonist cycles. The IVM protocol, FSH and hCG priming with oocyte retrieval 38 hours later, is an effective ART option that is comparable with conventional IVF for infertile women with PCOS.

• Clinical and Experimental Reproductive Medicine
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• 제23권3호
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• pp.303-310
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• 1996

본 연구는 epidermal growth factor (EGF)가 처리된 돼지 체외성숙란의 채외수정 후 배발달을 조사하기 위하여 실시하였다. 난구세포가 치밀한 미성숙란은 TCM 199 배양액에 (1) 무처리군 (2) 10 ng/ml EGF 처리군 (3) 10 ${\mu}g/ml$ FSH와 10% FBS 처리군 (4) 10 ng/ml EGF와 10 ${\mu}g/ml$ FSH 그리고 10% FBS 처리군으로 나누어 42시간 동안 배양하였다. 실험 1은 이들 처리군을 수정 후 NCSU (0.4% BSA) 배양액에서 배양하여 후기 배발달을 조사하였다. 그 결과 (3)과 (4)처리군 (13.4, 18.3%)의 배반포로의 발달율이 (2) 처리군(5.2%, p < 0.005)보다 현저하게 높았지만, (2) 처리군의 경우 (1) 처리군(1.2%, p < 0.005)의 배반포로의 발달보다는 현저하게 높았다. 실험 2는 수정 이후 6일째 이들 처리군에서 생산된 배반포기배를 double staining (propidium iodide and bisbenzimide) 방법을 이용하여 세포수를 조사하였다. 그 결과 (4) 처리군 (58.80 ${\pm}$ 11.90)의 배반포의 total 세포수가 (2)와 (3) 처리군 (42.17 ${\pm}$ 9.97, 49.07 ${\pm}$ 9.77, p < 0.05)보다 현저하게 많았으며, 배반포의 ICM 수에 있어서도 (4) 처리군 (11.69 ${\pm}$ 5.56)이 (2)와 (3) 처리군 (5.00 ${\pm}$ 4.24, 6.77 ${\pm}$ 4.92, p < 0.05)보다 현저하게 많았다. 더 우기 (4) 처리군 (19.0 ${\pm}$ 1.6)의 total 세포수에 대한 ICM의 비율이 (2)와 (3) 처리군 (11.1 ${\pm}$ 3.0, 12.7 ${\pm}$ 2.1) 보다 높았다. 이들의 결과로서, EGF가 단독 처리된 돼지 체외성숙란은 비록 낮았지만 배반포로 발달할 수는 있으나, 체외성숙시 FSH와 FBS의 추가 첨가는 높은 배반포로의 발달과 total 세포수와 ICM의 증가를 가져올 수 있음을 알수 있었다.

• Clinical and Experimental Reproductive Medicine
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• 제33권3호
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• pp.189-198
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• 2006

목적: 본 연구진은 난자성숙 과정의 조절 기작을 규명하기 위하여 생쥐의 미성숙 난자와 성숙난자에서 차이 나게 발현하는 유전자의 목록을 얻은 바 있다. 이들 유전자 중에서 Bcl-2 homolog인 Diva 유전자가 난자에 특이적으로 발현함을 본 연구를 통해 규명하였는데 이러한 Diva의 기능을 밝혀내기 위하여 immunoprecipitation (IP)과 Mass Spectrometry (MS)를 이용하여 Diva와 결합하여 상호작용하는 단백질을 동정하고자 하였다. 연구방법: NIH/3T3 세포주에 Diva를 encoding하는 pCMV-FLAG-Diva를 24 시간 동안 과발현 시키고 대조군으로는 유전자 없는 pCMV-FLAG empty vector를 transfection 하였다. FLAG에 특이적인 항체로 IP하여 Diva와 결합하는 면역복합체를 형성하게 한 후 이를 12% SDS-polyacrylamide gel 상에서 전기 영동하였고 Coomassie Blue 염색을 통해 단백질 발현양상을 관찰하였다. 대조군에서는 관찰되지 않으면서 Diva 유전자가 발현하는 실험군에서만 확인되는 밴드를 오려내어 trypsin을 사용하여 in-gel digestion 한 후 MS 분석을 시행하였다. 모든 mass spectra는 4700 Proteomics Analyzer (Applied Biosystems, Framingham, MA)에 의해 positive reflector mode에서 얻어졌다. 이렇게 얻어진 단백질들은 MASCOT Peptide Mass Fingerprint software (Matrixscience, London)을 이용하여 NCBI nonredundant database를 찾아서 동정하였다. 결과: Diva를 과발현하는 세포주에서만 관찰되는 15개 밴드에 대한 MS/MS 분석 결과, Diva와 결합하는 단백질로서 actin과 그 외에 ${\alpha}$-actinin, tropomyosin, tropomodulin 3 등의 actin-binding 단백질을 동정하였다. Diva를 과발현하는 NIH/3T3 세포주에서 면역 복합체를 형성하는 actin과 tropomyosin이 실제 난소 조직에서도 Diva와 결합하는지 IP와 Western blot을 통해 확인한 결과, actin과 tropomyosin 모두 Diva와 결합함을 확인함으로써, Diva는 이 두 단백질과 난소에서 상호작용함을 알 수 있었다. 결론: 본 연구는 Diva와 결합하여 상호작용하는 단백질들이 cytoskeletal system의 actin filament와 관계 있음을 규명한 최초의 보고이다. Diva가 actin과 tropomyosin과 결합하는 것을 고려해볼때, 난자 특이적인 Diva는 아마도 난자성숙 동안에 cytoskeletal system 을 조절하는 역할을 할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권12호
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• pp.1820-1826
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• 2007

In this study, we conducted various experiments in order to develop enhanced cultural conditions for in vitro-produced porcine embryos. All embryos were produced by in vitro maturation (IVM) and fertilization (IVF) of immature oocytes from abattoir-derived ovaries. In experiment 1, we cultured IVF embryos in 4 different groups, namely, 0% bovine serum albumin (BSA), 3% BSA, 0.05% Polyvinyl alcohol (PVA), and 0.5% Polyvinylpyrrolidone (PVP) added to the basal fluid cultural medium, Porcine zygote medium 3 (PZM-3). The rates of embryo development were higher in the group where the PZM-3 media had been supplemented with 3% BSA than the other groups. While not statistically significant, the percent of blastocysts and hatched blastocytes were 6.9% and 25.0% in the 3% BSA group vs. 1.2-6.4% and 0-16.7% in the other groups, respectively. In experiment 2, we added 10% fetal bovine serum (FBS) to PZM-3 on day 0 of culture and observed the development rate of blastocysts per day of culture from days 0 to 5. The development rate of blastocysts was higher at 15.6% on day 4 than on any other day, and was significantly higher than on day 0 or day 1 (p<0.05). The development rate of hatched blastocysts was 26.7% on day 4, and was higher than on any other day. In experiment 3, we cultured IVF embryos with different fluid culture media, grouped as 1) PZM-3+0.3% BSA (day0-day7); 2) PZM-3+0.3% BSA${\rightarrow}$day-4) PZM-3+10% FBS; 3) PZM-3+0.3% BSA${\rightarrow}$PZM-3+0.3% BSA+(day-4) FBS 10%; and 4) PZM-3+0.3% BSA+10% FBS (day0-day7). The development rates of blastocysts and hatched blastocysts were 21.5% and 53.1% in group 3, respectively, which was significantly higher than group 4 with respect to blastocyst development (5.2%, p<0.05) but not hatched blastocysts (14.3%). The total cell number (TCN) of blastocysts in group 3 was higher at $37.8{\pm}16.1$ than the other groups at $16.8{\pm}4.4$ - $30.1{\pm}10.9$; however, this was not significantly different. The results of this study showed that PZM-3 containing 0.3% BSA and supplemented with FBS during the later stage of culture on day 4 resulted in better TCNs and an increased rate of hatched blastocysts.

• 한국수산과학회지
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• 제45권3호
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• pp.253-261
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• 2012

We investigated the reproductive capacity of starry flounder Platichthys stellatus from the Uljin (UJ) marine ranching area and compared it to that of P. stellatus from the coastal waters of Pohang (PH). In UJ, female gonadosomatic index (GSI) peaked in October ($3.14{\pm}0.87$) and male GSI was high in October and December. In PH, female GSI peaked in January ($18.64{\pm}2.15$) while male GSI began to increase in October and remained high until March. Most ovaries of UJ females were immature with perinucleus oocytes, although the testes of UJ males were ripe in January. Both the ovaries and testes of PH starry flounders were ripe from January to March. The plasma estradiol-$17{\beta}$ ($E_2$) levels of UJ females were highest in October ($4.09{\pm}1.90$ ng/mL) although the testosterone (T) levels of UJ males were highest in December ($3.81{\pm}0.78$ ng/mL) and decreased gradually until April. The $17{\alpha}$,$20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) levels of UJ females were not detected. The E2 levels of PH females were highest in December ($36.25{\pm}33.07$ ng/mL) and $17{\alpha}20{\beta}P$ levels were highest in March ($5.51{\pm}0.95$5 ng/mL). The T levels of PH males were highest in December ($4.03{\pm}1.34$ ng/mL) and decreased gradually until October. Taken together, these results suggest that most females from UJ did not reach maturation with a spawning period that was considered to be between December and January.