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人类卵母细胞的激光辅助胞浆内单精子注射

Laser-assisted intracytoplasmic sperm injection in human oocytes.

作者信息

Eroglu Ali, Nahum Ravit T, Isaacson Keith, Toth Thomas L

机构信息

Vincent Memorial Obstetrics and Gynecology Service, Massachusetts General Hospital and Harvard Medical School, and Surgical Services, Massachusetts General Hospital, Harvard Medical School and Shriners Burns Hospital, Boston, Massachusetts, USA.

出版信息

J Reprod Med. 2002 Mar;47(3):199-203.

Abstract

OBJECTIVE

To explore potential applications of a non-contact, 1,480-nm diode laser to intracytoplasmic sperm injection (ICSI) of human oocytes.

STUDY DESIGN

Human oocytes obtained from in vitro fertilization (IVF) patients and failed to fertilize 24-48 hours after conventional IVF were used for ICSI along with discarded sperm. A noncontact, 1,480-nm diode laser was employed to immobilize sperm, to open a hole in the zona pellucida (ZP) and to perform ICSI through the hole. After ICSI and its simulation, oocytes were examined for formation of pronuclei, cleavage and normality of the cytoskeleton.

RESULTS

The 1,480-nm diode laser permitted fast and easy sperm immobilization and microdrilling of ZP to facilitate microinjection. Of the 78 injected oocytes, 53 (68%) survived the procedure, and 13 (25%) of them formed two pronuclei by 18 hours. Further culture of two fertilized eggs resulted in cleavage up to the eight-cell stage before cease of culture. Four oocytes were fixed after simulation of the procedure without injecting sperm. None of them showed gross abnormalities in cytoskeletal organization.

CONCLUSION

A noncontact, 1,480-nm diode laser can be used for the immobilization of sperm and for opening a hole in the ZP to facilitate ICSI, biopsy manipulation toward preimplantation genetic diagnosis and assisted hatching.

摘要

目的

探讨非接触式1480纳米二极管激光在人类卵母细胞胞浆内单精子注射(ICSI)中的潜在应用。

研究设计

从体外受精(IVF)患者获取的人类卵母细胞,在常规IVF后24 - 48小时未受精,将其与废弃精子一起用于ICSI。使用非接触式1480纳米二极管激光固定精子,在透明带(ZP)上开孔,并通过该孔进行ICSI。在ICSI及其模拟操作后,检查卵母细胞原核的形成、卵裂及细胞骨架的正常情况。

结果

1480纳米二极管激光能够快速简便地固定精子并对ZP进行微钻孔以利于显微注射。在78个注射的卵母细胞中,53个(68%)在操作后存活,其中13个(25%)在18小时时形成了两个原核。两个受精卵进一步培养至八细胞阶段后停止培养。在模拟操作但未注射精子后对4个卵母细胞进行固定。它们均未显示细胞骨架组织有明显异常。

结论

非接触式1480纳米二极管激光可用于固定精子以及在ZP上开孔,以利于ICSI、针对植入前基因诊断的活检操作和辅助孵化。

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