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대한생식의학회지 제26권 제2호 2010년
생쥐 수정란의 보조부화술에 있어서 non-contact type인 diode laser의 이용
을지병원 의과학연구소1, 산부인과2, 대전을지대학병원 산부인과3, 건국대학교 축산학과4
김동훈1, 이명섭3, 강희규1, 한성원1, 김묘경1, 박원일2, 이훈택4, 정길생4, 이호준1,
Use of Non-Contact Type Diode Laser on Assisted Hatching of Mouse Embryos
Kim dh, Lee ms, Kang hg, Han sw, Kim mk, Park wi, Lee ht, Chung ks, Lee hj
The present study was performed to investigate the efficacy and safety of laser assisted hatching (AH) on mouse embryos. Non-contact 1.48 ㎛ 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 was 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 ㎛ 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.
키워드 : laser assisted hatching, 1.48 ㎛ diode laser, anti-hatching system
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