생쥐 수정란의 보조부화술에 있어서 Non-Contact Type인 Diode Laser의 이용

Use of Non-Contact Type Diode Laser on Assisted Hatching of Mouse Embryos

  • Kim, D.H. (Medical Science Institute of Eulji General Hospital) ;
  • Lee, M.S. (OB/GYN of Eulji General Hosptial) ;
  • Kang, H.G. (Medical Science Institute of Eulji General Hospital) ;
  • Han, S.W. (Medical Science Institute of Eulji General Hospital) ;
  • Kim, M.K. (Medical Science Institute of Eulji General Hospital) ;
  • Park, W.I. (OB/GYN of Eulji Medical College Hospital) ;
  • Lee, H.T. (Department of Animal Science, Kon-Kuk University) ;
  • Chung, K.S. (Department of Animal Science, Kon-Kuk University) ;
  • Lee, H.J. (Medical Science Institute of Eulji General Hospital)
  • 발행 : 1999.06.30

초록

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.

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