• Clinical and Experimental Reproductive Medicine
• /
• 제35권4호
• /
• pp.275-283
• /
• 2008

목 적: 본 연구는 유리화동결 전 인공수축과 보조부화술 (부분투명대절개)이 유리화동결 후 생쥐 포배아의 생존율과 부화율에 미치는 영향을 알아보기 위해 시행되었다. 연구방법: 생쥐 2-세포기 배아를 획득하여 G1.1과 G2.2 배양액에서 포배아까지 배양하였다. 실험은 대조구군과 3개의 처리군으로 나누었다. 인공수축과 보조부화술 없이 동결한 군 (-AS/-AH)을 대조군으로 하였고, 보조부화술만 시행한 군 (-AS/+AH), 인공수축만 시행한 군 (+AS/-AH), 인공수축과 보조부화술을 동시에 처리한 군 (+AS/+AH)을 처리군으로 하였다. 모든 포배아는 G10과 G10E20 용액에서 각각 3분씩 평형을 실시하였고, G25E25 유리화용액에 노출직후 capped pulled-straw에 mouth 피펫으로 부하하여 유리화동결하였다. 융해 후 24시간 동안 배양하면서 생존율과 부화율을 조사하였다. 결 과: 인공수축을 시행한 후 보조부화술을 시행한 군 (+AS/+AH)과 보조부화술을 시행하지 않은 군 (+AS/-AH)에서의 생존율과 부화율은 각각 98%와 100%, 92%와 41%였으며, 인공수축을 시행하지 않고 보조부화술을 시행한 군(-AS/+AH)과 보조부화술을 시행하지 않은 군 (-AS/-AH, 대조군)에서의 생존율과 부화율은 각각 54%와 96%, 58%와 34%를 나타내어 인공수축과 보조부화술 생쥐 포배아의 생존율과 부화율 향상에 매우 효과적인 방법임을 알 수 있었다 (p<0.01). 또한 capped pulled-straw라고 명명한 스트로우를 이용한 유리화동결은 융해 후 회수율이 100%였으며, 동결과 융해과정에 매우 편리하여 배아의 유리화동결에 매우 유용할 것으로 생각된다. 결 론: 인공수축과 보조부화술은 포배아의 유리화동결 후 생존율과 부화율을 유의하게 향상시켜 포배아의 동결보존에 매우 효과적인 방법으로 생각된다.

• Clinical and Experimental Reproductive Medicine
• /
• 제35권3호
• /
• pp.193-202
• /
• 2008

목 적: 보조부화술이 적용되는 좋지 않은 예후를 보여주는 선별된 환자군을 대상으로 산성 용액을 이용한 AHA 방법과 레이저를 이용한 AHL 방법의 효용성을 비교하여, 보다 효과적으로 임신율과 착상률을 높일 수 있는 보조부화술 방법을 찾고자 하였다. 연구방법: 2006년 2월부터 9월까지 체외수정 시술을 시행한 환자 중 보조부화술이 필요한 328주기를 대상으로 산성용액을 이용한 AHA 방법 (180주기)과 ZILOS-tk 레이저를 이용한 AHL 방법 (148주기)으로 나누어 시행하였다. 보조부화술을 시행한 환자군은 환자의 나이가 38세 이상이거나 투명대의 두께가 $18{\mu}m$ 이상, 기저 FSH 농도가 15 mIU/ml이상, 체외수정 시술을 3번 이상 실패한 환자, 이식하는 배아의 상태가 양호하지 않은 환자들로 이상에 적용요인이 있으면 시행하여 무작위로 보조부화술 방법간에 환자들의 임상적 특징과 임신율과 착상률을 분석하였다. 결 과: 전체 보조부화술을 시행한 환자군에 AHL 방법과 AHA 방법간에 임신율 (42.6%, 63/148 vs. 33.3%, 60/180)과 착상률 (17.4%, 82/470 vs. 16.0%, 89/556)에 유의적 차이는 나타나지 않았다. 그러나 나이가 많은 환자군인 Group 1은 임신율 (37.0%, 20/54 vs. 18.7%, 14/75)과 착상률 (14.4%, 23/160 vs. 7.1%, 15/210)이 AHL 방법이 AHA 방법보다 유의적 (p<0.05)으로 높게 나타났다. 전체 환자군이나 선별된 각 군내에 보조부화술 방법간에 환자의 임상적 특징은 유의적 차이가 나타나지 않았다. 3번 이상 체외시술에 실패한 환자군 [Group 2: 43.8% (21/48)과 31.6% (25/79)], 투명대의 두께가 $18{\mu}m$ 이상인 환자군 [Group 3: 43.8% (32/73)과 34.1% (28/82)], 이식한 배아의 질이 양호하지 않은 환자군[Group 4: 25.0% (7/28)과 14.6% (6/41)]에서는 AHL 방법이 임상결과는 좋았으나 유의적 차이는 없었다. 결 론: 레이저를 이용한 AHL 방법이 나이가 많은 환자군과 3번 이상 체외수정 시술에 실패한 환자군에서 AHA방법에 비해 높은 임신율과 착상률을 나타내었다. 결론적으로, AHL을 이용한 보조부화술이 임상적으로 보다 효과적이고 안전한 방법이라고 사료된다.

• 대한생식의학회:학술대회논문집
• /
• 대한불임학회 1997년도 제34차 추계 학술대회
• /
• pp.64.1-64.1
• /
• 1997

• 대한생식의학회:학술대회논문집
• /
• 대한불임학회 1995년도 추계 학술대회 및 정기총회
• /
• pp.57-58
• /
• 1995

• 대한생식의학회:학술대회논문집
• /
• 대한불임학회 2002년도 제43차 추계학술대회
• /
• pp.88-89
• /
• 2002

• 대한생식의학회:학술대회논문집
• /
• 대한불임학회 1999년도 제38차 추계 학술대회
• /
• pp.110.2-111
• /
• 1999

• Clinical and Experimental Reproductive Medicine
• /
• 제22권2호
• /
• pp.183-189
• /
• 1995

In human IVF-ET, the development and morphology of the embryo have been known to affect implantation and pregnancy rates(PRs). Recently, pregnancy has been reported to related to the embryos with thick zona-pellucida, high levels of fragmentation, poor blastomere development and zona hardening. Although the mechanism of implantation is unclear, it is thought that the hatching process precedes implantation and that the hatching is related to implantation and PRs. This study was carried out to investigate the effect of assisted hatching(AHA) on the improvement of PRs in human IVF-ET. The results were as follows; 1. The PRs of the AHA group (40.8%) was significantly higher than that of control group(27.2%)(P<0.01). 2. According to the age of patients, the PRs of control and AHA groups were 33.9%(20/59), 44,4%(12/27) in <30 yrs, 26.1%(30/115), 38.3%(18/47) in 31-35 yrs, 22.4%(13/58), 41.4%(12/29) in >36 yrs, respectively. 3. According to the factors of infertility in AHA group, unexplained(immunologic factor) (40.0%) and male factors(41.9%) were higher than female(tubal obstruction, endometriosis, adhesion) factor (28.9%). As a result, it is suggested that AHA technique improve the PRs in poor prognosis patients. It is concluded that AHA method can be used to improve the PRs in human lVF-ET.

• Clinical and Experimental Reproductive Medicine
• /
• 제28권2호
• /
• pp.147-153
• /
• 2001

Objective: The present study was performed to investigate the efficiency of partial laser assisted hatching (p-LAH; lased 1/2 ZP width from ZP edge) on hatching of mouse blastocysts. Methods: We used non-contact $1.48{\mu}m$ diode laser (MTM, Switzland) to create a precise hole on zona pellucida. 2-cell embryos were collected from the mouse (ICR) oviduct at 48 hours after hCG administration. Collected 2-cell embryos were cultured in the P-1 medium supplemented with 0.4% BSA. For experiments, embryos at 8-cell stage were used after $20{\sim}22$ hours in culture. After conventional (c-LAH) or partial laser assisted hatching, the embryos were further cultured in P-1 medium supplemented with 0.4% BSA for 3 days. To compare efficiency of complete and partial laser assisted hatching, hatching rate, hatching time and blastocyst diameter and zona pellucida thickness at hatching time were investigated. Embryos were examined every 12 hours. Blastocyst diameter and zona pellucida thickness at hatching time were measured with an ocular micrometer. Results: Hatching rates of p-LAH group (84.2%) was significantly higher than that of control group (39.3%), but there was no difference between the p-LAH (84.2%) and c-LAH (91.2%). p-LAH group was hatched 12 hours earlier than control group, but hatched 12 hours later than c-LAH group. The diameter of blastocyst at hatching time of p-LAH group ($113.1{\pm}6.4{\mu}m$) was smaller than that of control group ($122.2{\pm}5.0{\mu}m$), but larger than that of c-LAH group ($102.2{\pm}2.7{\mu}m$). Zona pellucida thickness at hatching time of p-LAH group ($6.4{\pm}0.9{\mu}m$) was thicker than that of control group ($4.5{\pm}1.5{\mu}m$), but thinner than that of c-LAH group ($10.0{\pm}0.8{\mu}m$). Conclusion: These results suggest that p-LAH may maintains the cell arrangement of early embryos to ensure successful development and prevent precocious hatching of blastocyst when compare to c-LAH and conventional (acidic tyrode) AH. Thus, p-LAH may provide a valuable and effective AH technique for human ART program.

• Clinical and Experimental Reproductive Medicine
• /
• 제25권2호
• /
• pp.123-128
• /
• 1998

Implantation rates remain low following human in vitro fertilization (IVF). Suboptimal culture conditions may limit the ability of embryos to hatch as blastocysts, and artificial opening of the zona pellucida has been proposed as a means to promote subsequent hatching (assisted hatching). In this study, assisted hatching (AH) by zona drilling using acidic Tyrode's solution was performed in 320 patients, due to their age of more than 38 years (group A), the thick zona pellucida (group Z; $ZP\geq0.18{\mu}m$), and failures in implantation more than 3 times in previous IVF-ET trial (group P). This study was designed firstly, to study the effects of AH on the outcomes of IVF-ET according to the indications and secondly, to verify the appropriate application of AH. The results were as follows; 1. There was no difference in pregnancy rate between AH group (26.6%) and non-AH group (26.5%). 2. Assisted hatching (AH) showed significantly higher pregnancy rate of the patients with thick zona pellucid a than those of the patients with age factor and with the history of repeated implantation failure. But in the patients with age factor only, AH resulted in higher pregnancy rate. 3. Interestingly, the patients with complex factors including zona factor (Z: 33.9%; ZA: 30.4%; ZP: 31.6%; ZAP: 21.4%) showed higher pregnancy rates than other complex factors excluding zona factor (A: 24.4%, P: 0%; AP: 10.8%). From these results, AH is more helpful to the patients with thick zona pellucida rather than patients with older age and/or previous repeated implantation failure.

• Clinical and Experimental Reproductive Medicine
• /
• 제26권2호
• /
• pp.185-192
• /
• 1999

The present study was performed to investigate the efficacy and safety of laser assisted hatching (AH) on mouse embryos. Non-contact $1.48{\mu}m$ diode laser system used to create a precise hole on zona pellucida. 2-cell embryos were collected from the mice (ICR) that had the coitus vaginal plug confirmed at 48 hours after hCG injection. Collected 2-cell embryos were cultured in the HTF medium supplemented with 0.4% BSA. For experiments, embryos at 8-cell stage were used after 18-22 hours in culture. After assisted hatching, the embryos were further cultured in HTF medium containing 0.1% PVP (anti-hatching system) for 3 days. For evaluate efficiency of laser on mouse embryo hatching, the effect of AH methods (acidic tyrode, pronase and laser), the number of artificial holes (1, 2 and 3 hole) and the irradiation time of laser (2, 4, 6, 8 and 10 ms) were examined. Hatching rates of laser AH group (95.2%) was significantly higher than that of control group (50.8%), but there was no differences among the laser (95.2%), acidic tyrode (100%) and pronase (98.5%) groups. Hatching rates of the number of zona pellucida opening by laser, there were no differences among the 1 hole (87.5%),2 hole (92.1%) and 3 hole (85.9%) groups. Developmental and hatching rates of embryos according to laser irradiation time were similar in the treatment groups. Therefore, these results suggest that laser AH using non-contact $1.48{\mu}m$ diode laser is a simple and accurate and effective procedure for AH. Based on these results, laser AH could be use safely for human ART program.