• Title/Summary/Keyword: Human IVF

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Identification and Characterization of Bull Sperm Hyaluronidase Hyal5

  • Kim, Ekyune;Chang, Kyu-Tae
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.491-498
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    • 2011
  • During mammalian fertilization, germ cell-specific hyaluronidases, such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5), are important for the dispersal of the cumulus mass. In this study, we demonstrated that bull Hyal5 is a single copy gene on chromosome 4 that is expressed specifically in the testis. In addition, we expressed recombinant bull SPAM1 and Hyal5 in human embryonic kidney 293T cells and showed that these enzymes possessed hyaluronidase activity. We also demonstrated that a polyclonal antibody against bull sperm hyaluronidase inhibits sperm-egg interactions in an in vitro fertilization (IVF) assay. Our results suggested that bull Hyal5 may have a critical role in bull fertilization.

Clinical Application of Oocyte Cryopreservation I. Pregnancy and Delivery of Vitrified Human Oocytes in ART Program (난자동결보존의 임상적 응용 I. 유리화 난자동결 보존에 의한 임신과 분만)

  • 정형민;박이석;차광렬
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.245-250
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    • 2001
  • This study was performed to evaluate whether vitrification method using ethyle glycol and eletron microscopic (EM) grid could be used far the cryopreservation of human oocytes in ART program. Surplus oocytes were obtained from consented IVF patients. These surplus human oocytes were frozen with our vitrification method, Oocytes were exposed to 1.5M ethylene glycol (EG) in DPBS far 2,5 minutes, followed by 5.5M EG plus 1.0M Sucrose in DPBS for 20 seconds. Then oocytes were transferred onto the EM grid and the grid was plunged into LN2 for storage. For thawing, oocytes containing EM grid were sequentially transferred in 1.0M, 0.5M, 0.25M, 0.125M and 0 M sucrose in DPBS solution at the intervals of 2.5 minutes. Thawed and survived oocytes were provided for ICSI. Embryos from vitrified oocytes were transferred to uterus of the patient on 4 to 5 days after ovulation in natural cycles of on 15 to 17 day of hormone replacement cycles. A total of 370 oocytes from 26 patients were thawed and 159 (43.0%) of them survived. One hundred thirty four oocytes (84.3%) were fertilized normally and 126 pre-embryos were transferred to 26 patients, resulting in 5 clinical pregnancies. The pregnancy rate per transfer was 19.2% and implantation rate was 4.0%. Among the five pregnant, 4 patients delivered 4 healthy babies and the one patient was 32-week ongoing pregnancy. From this results, vitrification using ethylene glycol as cryoprotectant and EM grid is a rapid and simple method that can be effectively applied for the cryopreservation of human oocytes in ART program.

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Influence of Human Follicular Fluid for IVF on PN Grade and Development of Human Embryos (사람 난포액에 의한 정자 처리가 체외수정란의 전핵과 발달에 미치는 영향)

  • B. G. Jeon;Lee, S. L.;S. A. Ock;Kim, K. S.;J. S. Moon;D. O. Kwack;Park, G. J.;S. Y. Choe
    • Journal of Embryo Transfer
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    • v.18 no.2
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    • pp.125-134
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    • 2003
  • 본 연구는 사람의 시험관아기 프로그램에서 체외 수정란의 질을 향상시키는 한 방법으로 난포액으로 처리된 정자를 체외 수정에 사용하여 생산된 체외 수정란의 전핵 등급과 발달 능력을 조사하였다. 정상적인 시험관 아기 시술을 시행한 실례 환자를 대상으로 과배란을 유기하여 배란 직전의 난자를 채취하여, 난포액으로 처리된 정자와 체외 수정시킨 후 사람 체외 수정란의 전핵 등급과 체외발달율을 조사하였다. 체외수정을 위한 정자의 처리 방법으로 synthetic serum substitute (SSS)를 15% 첨가한 modified human total fluid (m-hTF) 혹은 난포액에서 정자를 2시간 동안 swimming-up 처리 후 각각 체외수정에 사용한 결과, 수정율은 75.3 및 82.1%를 나타내어 유의적인 차이는 없었으나, 1등급 전핵란은 각각 48.0 및 65.5%를 나타내어 난포액에서 유의적으로 (P<0.05) 높았고, 배양 후 3일째에 수정란의 등급을 조사한 결과, 1등급 체외수정란은 각각 44.9 및 60.5%를 나타내어 난포액에서 유의적으로 (P<0.05) 높았다. 또한 체외수정란을 배양 후 3일째에 수정란의 발달 단계를 조사 한 결과, 5-세포기 단계 이상을 발달하는 비율은 각각 51.0 및 70.5%를 나타내어 난포액에서 유의적으로 (P<0.05) 높았다.

Increased cryo-survival rate in ejaculated human sperm from infertile men following pre-freeze in vitro myo-inositol supplementation

  • Saleh, Ramadan;Assaf, Hanan;Abd El Maged, Wafaa M.;Elsuity, Mohamed;Fawzy, Mohamed
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.4
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    • pp.177-182
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    • 2018
  • Objective: To investigate the effects of in vitro myo-inositol (Myo-Ins) supplementation of cryopreserved human semen on the cryo-survival rate (CSR). Methods: Semen samples were obtained from 41 infertile men. Following routine semen analysis, each sample was divided into two equal aliquots (0.5 mL each). One aliquot was treated with 1 mg of Myo-Ins dissolved in $10{\mu}L$ of sperm preparation medium. The second aliquot was treated with $10{\mu}L$ of the same medium (control). Both aliquots were incubated for 20 minutes prior to freezing to slow the freezing process. The frozen samples were examined for post-thaw percentages of total motility (TM), progressive motility (PM), and the CSR, defined as the percentage of post-thaw TM divided by the percentage of pre-freeze TM and multiplied in 100. The results were expressed as median and interquartile range (25th and 75th percentiles). Results: The pre-freeze TM (50% [30%-50%]) and PM (35% [20%-35%]) were significantly higher than the post-thaw TM and PM in the MyoIns group (15% [10%-35%] and 10% [5%-20%]; p<0.001 and p<0.001, respectively) and the control group (10% [6%-30%] and 5% [3%-15%]; p<0.001 and p<0.001, respectively). The CSR of the 41 semen aliquots supplemented with Myo-Ins (40% [25%-70%]) was significantly higher than that of the control samples (30% [13%-58%], p=0.041). The CSR of the 26 abnormal semen samples that were supplemented with Myo-Ins (38% [20%-50%]) was significantly higher than that of the control samples (23% [12%-30%], p=0.031). Conclusion: In vitro Myo-Ins supplementation of ejaculated human sperm from infertile men resulted in a significant increase in the CSR in samples with abnormal pre-freeze sperm parameters.

Clomid/hMG/hCG Regimen or FSH/hMG/hCG Regimen : Differences in The Luteal Phase Serum Estradiol and Progesterone Levels Determined after In Vitro Fertilization (시험관아기 프로그램에서 과배란유도 약제의 종류에 따른 황체기 혈청 난포호르몬 및 황체호르몬의 변동에 관한 비교연구)

  • Chang, Yoon-Seok;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Lee, Jin-Yong;Kim, Hak-Soon
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.1
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    • pp.69-79
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    • 1989
  • Steroid hormone profiles during luteal phase of clomiphene citrate(CC)/human menopausal gonadotropin(hMG)/human chorionic gonadotropin(hCG)-stimulated in vitro fertilization (IVF) cycles and of follicle-stimulating hormone(FSH)/hMG/hCG-stimulated IVF cycles were compared. In seventy three cycles stimulated with CC/hMG/hCG regimen, follicles were aspirated during exploratory laparotomy and yielded 7 pregnancies, and in 83 cycles stimulated with FSH/hMG/hCG regimen, follicles were aspirated by laparoscope and made 13 pregnancies. Serum estradiol($E_2$) and progesterone($P_4$) levels were determined on days 2, 5, 7, and 9 after follicle aspiration. The FSH/hMG/hCG regimen was more effective than the CC/hMG/hCG regimen in folliculogenesis, ie, ovarian stimulation, follicular phase $E_2$ peak levels, oocyte maturation, and the number of retrieved oocytes. There was no significant difference between luteal serum $P_4/E_2$ ratio of the two regimens, suggesting that secretory endometrial build-up ability for implantation may not differ each other. Several significant correlations were observed between follicular phase seum $E_2$ peak levels and luteal phase serum $E_2$ and $P_4$ levels in the FSH/hMG/hCG-stimulated cycles but any correlation was not significant in the CC/hMG/hCG-stimulated cycles, suggesting that somewhat more follicles may eventually fall in atresia even after attaining dominant stage in the CC/hMG/hCG-stimulated cycles than the FSH/hMG/hCG-stimulated cycles.

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