• 한국수정란이식학회지
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• 제26권4호
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• pp.277-282
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• 2011

The present research was carried out to evaluate the possibility of increasing female offspring production ratios using artificial insemination buffer (AIB) before artificial insemination (AI). In this experiment, we optimized AIB composition, made an AIB gun and analyze factors affecting AI non-return rate after AIB treatment. The AIB was made with the base of Tris-buffer supplemented with L-arginine and several other chemicals that might reduce the motility of male sperm compared to the female counterpart, therefore, increasing the possibility of fertilization by female sperm. AIB must be deposited into $2^{nd}$ to $4^{th}$ cervix by AIB gun. After 15 min of AIB deposition, frozen semen was deposited into the same place. A total of 348 cattle were inseminated with AIB insemination, and there were no significant differences between AIB and traditional AI non-return rates (56.8% vs. 55.7%). The AI non-return rate in AIB group, however, differed significantly among 7 Hanwoo farms. The parturition numbers ($1^{st}$ to $7^{th}$) of cows did not affect AIB AI rate. The proportion of AIB AI success rates was significantly higher in Hanwoo cows than in dairy cows (61.0% vs. 48.7%), but the average AI success rate did not differ significantly between AIB and conventional AI (56.8% vs. 55.7%). The female offspring production rate in $2^{nd}$ to $4^{th}$ cervix deposition place was significantly higher than that in the uterus body (77.7% vs. 59.6%, p<0.05). The injection volume of AIB in 5 and 10 ml was significantly higher than that in 2 ml (77.7%, 78.7% vs. 51.8%, p<0.05), but there were no differences in AIB injection volume between 5 and 10 ml. The best exposure time of AIB in the cervix was 10 to 15 min rather than 5 min (79.2%, 77.2% vs. 52.6%, p<0.05). AIB therefore needs to have an exposure time of at least over 10 min for a higher production rate of female offspring. In conclusion, AIB could be used in AI industry to increase the female offspring ratio and AIB AI can increase the AI success rate.

• 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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• 제42권4호
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• pp.143-148
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• 2015

Objective: The aim of the current study was to determine the predictive value of anti-$M{\ddot{u}}llerian$ hormone (AMH) levels for pregnancy outcomes in patients over 40 years of age who underwent in vitro fertilization or intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycles. Methods: We retrospectively analyzed the medical records of 188 women aged 40 to 44 years who underwent IVF/ICSI-fresh ET cycles due to unexplained infertility in the fertility center of CHA Gangnam Medical Center. Patients were divided into group A, with AMH levels <1.0 ng/mL (n=97), and group B, with AMH levels ${\geq}1.0ng/mL$ (n=91). We compared the clinical pregnancy rate (CPR) in the two groups and performed logistic regression analysis to identify factors that had a significant effect on the CPR. Results: The CPR was significantly lower in group A than group B (7.2% vs. 24.2%, p<0.001). In multivariate logistic regression analysis, AMH levels were the only factor that had a significant impact on the CPR (odds ratio, 1.510; 95% confidence interval, 1.172-1.947). The area under the receiver operating characteristic curve for AMH levels as a predictor of the CPR was 0.721. When the cut-off level of AMH was set at 1.90 ng/ mL, the CPR was 6.731-fold higher in the group with AMH levels ${\geq}1.90ng/mL$ than in the group with AMH levels <1.90 ng/mL (p<0.001). Conclusion: Our study showed that AMH levels were predictive of clinical pregnancy in infertility patients over 40 years of age. Further prospective studies should be conducted to validate the predictive capability of AMH levels for the outcome of clinical pregnancy.

• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.33-39
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• 2017

Objective: The aim of this study was to assess the changes of follicular fluid (FF) and serum levels of cerebellin precursor protein 1 (cbln1) and betatrophin in patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with a gonadotropin-releasing hormone (GnRH) antagonist protocol. Methods: Twenty infertile women with PCOS and 20 control women diagnosed as poor responders undergoing ovarian stimulation with a GnRH antagonist were included. Blood samples were obtained during ovum pick-up. Follicular fluid from a dominant follicle was collected from the subjects. Using enzyme-linked immunosorbent assays, FF and serum levels of cbln1 and betatrophin were measured in both groups of participants. Metabolic and hormonal parameters were also determined and correlated with each other. Results: Both groups of women had similar serum and FF betatrophin levels ($55.0{\pm}8.9ng/mL$ vs. $53.1{\pm}10.3ng/mL$, p=0.11). The serum and FF betatrophin levels of poor responders were found to be similar ($49.9{\pm}5.9ng/mL$ vs. $48.9{\pm}10.7ng/mL$, p=0.22). Conversely, the FF cbln1 levels of PCOS women were found to be significantly higher than the serum cbln1 levels ($589.1{\pm}147.6ng/L$ vs. $531.7{\pm}74.3ng/L$, p<0.02). The FF cbln1 levels of control participants without PCOS were significantly higher than their serum cbln1 levels ($599.3{\pm}211.5ng/L$ vs. $525.3{\pm}87.0ng/L$, p=0.01). Positive correlations were detected among body mass index, insulin resistance, serum insulin, total testosterone, and betatrophin levels in the PCOS group. Conclusion: Follicular fluid betatrophin and cbln1 concentrations may play a pivotal role on follicular growth in PCOS subjects undergoing IVF/ICSI with an antagonist protocol.

• Clinical and Experimental Reproductive Medicine
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• 제34권1호
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• pp.41-48
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• 2007

목 적: 무정자증이나 심한 희소정자증이 있는 남성 불임 환자에서 Y 염색체 미세 결손을 동반하는 경우 체외 수정 및 세포질내 정자 주입술과 배아 이식의 결과에 미치는 영향을 알아보고자 하였다. 연구방법: 후향적 실험군 -대조군 연구로, Y 염색체 미세 결손이 동반된 무정자증이나 심한 희소정자증이 있는 남성 불임 부부 14 쌍을 실험군으로 하였고 대조군으로는 남성 배우자가 무정자증이나 심한 희소정자증이 있지만 Y 염색체는 정상인 12 쌍의 불임 부부를 대상으로 하였으며, 이 두 군을 다시 정자 채취 방법에 따라 사정된 정자를 이용한 군과 고환 내 정자 추출술을 통해 정자를 채취한 군으로 분류하였다. 연구 결과로는 수정률, 등급이 우수한 배아 수, 착상률, 융모막성 성선자극호르몬 양성률, 초기 임신 소실률, 출생률을 비교하였다. 결 과: 등급이 우수한 배아 수, 착상률, 융모막성 성선자극호르몬 양성률, 초기 임신 소실률, 출생률은 Y 염색체 미세 결손이 있는 군과 정상 Y 염색체를 갖는 군 간에 차이가 없었으나, 수정률은 Y 염색체 미세 결손이 있는 불임 부부 (61.1%) 에서 정상 Y 염색체를 갖는 불임 부부 (79.8%, p=0.003) 보다 낮았다. 정자 채취의 방법에 따라 비교하였을 때, 고환 내 정자 추출술을 통해 정자를 채취한 경우 실험군 (52.9%) 에서 대조군 (79.5%, p=0.008) 보다 유의하게 수정률이 낮게 나타났으나 사정된 정자를 이용한 경우에는 실험군에서 대조군 보다 수정률이 더 낮은 경향을 보였으나 유의한 차이는 없었고, 나머지 결과들은 두 군 간에 차이가 없었다. 결 론: Y 염색체 미세 결손이 동반된 무정자증이나 심한 희소정자증이 있는 불임 부부에서 고환 내 정자 추출술을 통해 정자를 채취할 경우 체외 수정 및 세포질내 정자 주입술 결과 정상 Y 염색체를 갖는 불임 부부에서 보다 수정률이 더 낮게 나타났으나, 사정된 정자를 이용하는 경우나 고환 내 정자 추출술을 통해 정자를 채취하여 수정이 된 후에는 Y 염색체의 미세 결손이 체외 수정 및 세포질내 정자 주입술과 배아 이식의 결과에 영향을 주지 않는 것으로 보인다. 그러나 향후 대규모의 전향적 연구가 더 필요할 것으로 사료된다.

• 한국가축번식학회지
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• 제20권1호
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• pp.53-61
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• 1996

This study was carried out to determine effectively the sex of rabbit embryos using H-Y antiserum. H-Y antiserum was obtained from inbred SD strain female rat which was immunized by injection of testis cell of inbred SD strain male rate into its spleen. The titer of antiserum was identified by sperm cytotoxicity test and culture of rabbit embryos with antiserum. The developed or undeveloped embryos were separated by exposure the embryos to the antiserum with H-Y antibody. Developed embryo were transferred to the recipients and sex of offspring were examined. 1. In the sperm cytotoxicity test, the rate of dead sperm showed no difference between two antisera from spleen and testis cell as antigens. It is confirmed that H-Y antibody in antiserum was absorbed by H-Y antigen in male rat spleen cells. 2. When rabbit morulae were exposed to antiserum and complement, the rate of embryos developed or arrested was 51 and 49% respectively and the rate was closely same as natural sex ratio of 50:50%. 3. When rabbit morulae were cultured for 12h in the medium containing antiserum produced by antigen of testis cell, the rate of embryos developed or arrested was 48 and 52% respectively and the rate was closely same as natural sex ratio of 50:50%. 4. Eighty rabbit embryos which were not affected by H-Y antiserum were transferred to four recipients. Two recipients were pregnant and born 13 pups among which 2 (14%) were male and 11 (86%) were female. In conclusion, existence of H-Y antibody in the serum from female rat immunized by injecting testis cell from newborn male rat to the spleen of the female rat was confirmed. When rabbitmorulae were exposed to H-Y antiserum and complement, about a half of embryos were developed to blastocysts. When the rabbit embryos not affected by H-Y antiserum were transferred, the rate of female offspring was 86%. Therefore, it was identified that most of embryos which were not affected by H-Y antiserum were female.

• Clinical and Experimental Reproductive Medicine
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• 제31권3호
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• pp.169-176
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• 2004

Objective: This study was performed to evaluate and compare the embryonic developmental capacity and pregnancy rates in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with ejaculated sperm or testicular sperm cycles. Materials and Methods: Fertilization was examined in the following morning after IVF (group I), ICSI (group II) or TESE-ICSI cycles (group III). Fertilized oocytes were co-cultured with Vero cells until embryo transfer (ET). On day 2 and $5{\sim}7$, grades of embryos (<4- or $\geq$4-cell) and blastocysts (BG1, 2, 3 or early) were evaluated. Clinical pregnancy rate was determined by detecting G-sac with transvaginal ultrasonogram. We analyzed the results by $X^2$ and Student's t-test and considered statistically significant when P value was less than 0.05. Results: Fertilization rate was significantly higher (p<0.05) in group I ($79.0{\pm}21.2%$) than in group II and III ($56.8{\pm}21.6%$ and $36.7{\pm}25.3%$). Cleavage and blastulation rate of group I ($95.8{\pm}13.8%$ and $59.5{\pm}25.3%$) were significantly higher (p<0.05) than those of group III ($83.4{\pm}18.6%$ and $40.4{\pm}36.5%$). Clinical pregnancy rate was significantly higher (p<0.05) in group I and II (40.7% and 41.7%) than that in group III (12.5%). No differences were found in the rates of multiple pregnancy and abortion among three groups. Embryonic implantation rate was higher in group I ($15.1{\pm}20.2%$, p<0.05) and II ($14.7{\pm}20.6%$, NS) than that in group III ($5.1{\pm}15.6%$). However, embryonic implantation rate was increased in ET with blastocyst(s) among three groups. Conclusions: Fertilized oocytes obtained from TESE-ICSI were harder to be successfully cultured to blastocyst stage for 5$\sim$7 days than that from IVF cycles. However, all blastocyst(s) ET increased the embryonic implantation rate equally in IVF, ICSI and TESE-ICSI cycles.

• 한국가축번식학회지
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• 제7권2호
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• pp.8-26
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• 1983

Various early pregnancy diagnostic methods have been developed in order to improve the reproductive efficiency in cow, mare, mule, sow, sheep, goat, dog, cat, rabbit, buffalo, camel, elephant, monkey, deer, lion, coipus and guinea pig. These methods include abdominal swelling, abdominal palpation, esturs cylce detection, Lupin test, gonadotropin assay, colostrum injection test, sperm motility assessment, cervical mucus viscosity test, Kaber chromagens method, estrogen test, A Scheim-Zond다 test, spectrophotometric detection of estrogen in urine and feces, boric acid crystraline formation test in urine, oxytocin injection test, diamino-oxidase test, PMSG HA test, behaviour test, Simolus iodine detection test, detection of tryptophane in urine, x-ray method, Cuboni and Lunaas method, vaginal biopsy method, Friedmann Schneider diagnostic method, electrode method, barium chloride detection method, ECG, Doptone method, ultrasound method, ultrasound scanning method, LDH method, rectal palpation method, CL palpation method, radioautography, serum creatine test, serum globulin test, chlormadine method, CAP method, Medata Do, pp.ers method, body fluid test, Plasma oCS detection method, ERIA, LHRH method, negative latex cogulation test and oestrone sulphate detection method. The most reliable methods with high a, pp.icability to farm animals such as sheep, mare, sow and cow are rectal palpation, ultrasound method and hormonal assay in blood and milk. However, they require complicated laboratory works for the early diagnosis of pregnancy and in most cases, the simple and economical methods which are described up to now need a long period of time after conception. Generally, it is possible to detect pregnancy after one estrus cycle, even though it varies depending on the species of animals. For improvement of the reproductive efficiency, it is required to develop a more accurate, economical, simple and early detectable method. It is anticipated that the result of a study on the detection method of EPF(early pregnancy factor) would be a, pp.icable to various animals within 6 hours after conception.

• 환경생물
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• 제38권2호
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• pp.323-331
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• 2020

해산송사리(Oryzias dancena)를 대상으로 한 발생공학 연구시, 현재의 자연산란법은 매우 수동적이고 불편하다. 본 연구에서는 적정 시간에 해산송사리의 수정란을 획득하고자 인공산란을 위한 인간융모성생식선자극 호르몬(human chorionic gonadotropin; HCG)과 잉어 뇌하수체(carp pituitary extract; CPE)의 최적 농도를 조사하였다. 정자활성도, 산란율, 수정 후 3일에서의 수정란 생존율 및 부화율은 100 HCG IU kg^-1 BW (body weight)와 CPE 5 mg L^-1에서 가장 높았다(p<0.05). 본 종에서의 수정란 필요시, HCG와 잉어 뇌하수체 주사를 통하여 적정시간에 미수정란과 정자를 획득할 수 있었다. 본 연구 결과는 해산송사리의 실험동물화 개발에 유용하리라 사료된다.

• 한국양식학회지
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• 제9권4호
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• pp.419-427
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• 1996

한해성 패류인 북방대합을 대상으로 자극방법별 산란유발 및 수온별 난발생 과정을 조사하였다. 모패에 대한 산란유발 자극은 $NH_4OH$ 첨가와 serotonin 주사가 효과적이었으며, 수컷이 암컷보다 민감하게 반응하였다. 반응개시 시간에 있어서 $NH_4OH$ 첨가시 방란은 $3\~4$시간, 방정은 1시간 전후에 개시되어 암수간의 차이가 있었다. serotonin 주사는 암수 모두에서 5분 이내에 방란${\cdot}$방정을 유도하였다. 방출된 알의 수는 $NH_4OH$ 첨가에서 유의하게 많았다. 그러나 serotonin 주사에 의해 산란유발된 알의 GVBD율, 수정률 및 부화율은 $NH_4OH$ 첨가에 의한 알보다 높았다. 방출된 알은 평균 난경 $65.2{\pm}1.8\;{\mu}m$인 분리 침성란으로 GVBD 이전에도 수정이 가능하였으며, 적정 발생수온은 $15\~20^{\circ}C$였다. 초기 D상 유생까지는 $15^{\circ}C$에서 42시간, $20^{\circ}C$에서 27시간이 소요되었다.