Successful Use of a Laser for Human Embryo Biopsy in Preimplantation Genetic Diagnosis: Report of Two Cases

Purpose:The use of Tyrode's acid to drill the zona pellucida for embryo biopsy is the most widely used methodology in preimplantation genetic diagnosis. Instead of this, we propose the use of a 1.48-μm diode noncontact laser, which is quicker, simpler, and safer.Methods:The laser beam was tangentially guided to the zona pellucida of the embryo. Depending on zona pellucida measurement, two to four consecutive shots of 8–22 msec were necessary to drill the zona pellucida of the 13 embryos biopsied for two patients (hemophilia carriers).Results:Female embryos were replaced into the uterus of the patients (1.5 embryos/replacement). One single pregnancy was established (33.3% implantation rate). Coculture of untransferable embryos showed a blastocyst rate of 66.7% (4/6) for male embryos and 25% (1/4) for abnormal ones.Conclusions:These results demonstrate the safety and usefulness of laser methodology in preimplantation genetic diagnosis.

[1]  B. Minhas,et al.  Survival and cell acquisition rates after preimplantation embryo biopsy: use of two mechanical techniques and two mouse strains. , 1990, American journal of obstetrics and gynecology.

[2]  J P Hornung,et al.  Improved fertilization and implantation rates after non-touch zona pellucida microdrilling of mouse oocytes with a 1.48 microm diode laser beam. , 1996, Human reproduction.

[3]  S. Beebe,et al.  Preclinical models for human pre-embryo biopsy and genetic diagnosis. I. Efficiency and normalcy of mouse pre-embryo development after different biopsy techniques. , 1992, Fertility and sterility.

[4]  M W Berns,et al.  MUTATION AND SISTER CHROMATID EXCHANGE INDUCTION IN CHINESE HAMSTER OVARY (CHO) CELLS BY PULSED EXCIMER LASER RADIATION AT 93 nm AND 308 nm AND CONTINUOUS UV RADIATION AT 254 nm , 1989, Photochemistry and photobiology.

[5]  H E Malter,et al.  Partial zona dissection of the human oocyte: a nontraumatic method using micromanipulation to assist zona pellucida penetration. , 1989, Fertility and sterility.

[6]  J. Gordon,et al.  Use of zona drilling for safe and effective biopsy of murine oocytes and embryos. , 1990, Biology of reproduction.

[7]  M W Berns,et al.  Micromanipulation of sperm by a laser generated optical trap. , 1989, Fertility and sterility.

[8]  A. Veiga,et al.  Cryopreservation of Blastocysts , 2001 .

[9]  M. Zilberstein,et al.  Preimplantation development of mouse and human embryos biopsied at cleavage stages using a modified displacement technique. , 1997, Human reproduction.

[10]  A. Handyside,et al.  Embryo biopsy strategies for preimplantation diagnosis. , 1993, Fertility and sterility.

[11]  J W Gordon,et al.  Clinical evaluation of three approaches to micromanipulation-assisted fertilization. , 1990, Fertility and sterility.

[12]  Z. Rosenwaks,et al.  Diagnosis of major chromosome aneuploidies in human preimplantation embryos. , 1993, Human reproduction.

[13]  A. Trounson,et al.  Biopsy of preimplantation mouse embryos: development of micromanipulated embryos and proliferation of single blastomeres in vitro. , 1989, Biology of reproduction.

[14]  J. Egozcue,et al.  The chromosome complement of first-cleavage mouse embryos after in vitro fertilization , 1986, Journal of in Vitro Fertilization and Embryo Transfer.

[15]  Jacques Cohen Assisted hatching of human embryos , 1991, Journal of in Vitro Fertilization and Embryo Transfer.

[16]  M. Berns,et al.  Opening of the mouse zona pellucida by laser without a micromanipulator. , 1993, Human reproduction.

[17]  A. Veiga,et al.  Laser blastocyst biopsy for preimplantation diagnosis in the human , 1997, Zygote.

[18]  P. Devroey,et al.  Efficiency and accuracy of polymerase-chain-reaction assay for cystic fibrosis allele ΔF508 in single cell , 1992, The Lancet.

[19]  Guy P. Delacretaz,et al.  1.48-um diode laser microdissection of the zona pellucida of mouse zygotes , 1994, SPIE LASE.

[20]  Klaus Rink,et al.  Non‐contact microdrilling of mouse zona pellucida with an objective‐delivered 1.48‐μm diode laser , 1996, Lasers in surgery and medicine.

[21]  A. Handyside,et al.  Human preimplantation development in vitro is not adversely affected by biopsy at the 8-cell stage. , 1990, Human reproduction.

[22]  A. Tarkowski An Air-Drying Method for Chromosome Preparations from Mouse Eggs , 1966 .

[23]  S Antinori,et al.  Seventeen live births after the use of an erbium-yytrium aluminum garnet laser in the treatment of male factor infertility. , 1994, Human reproduction.

[24]  M W Berns,et al.  Micromanipulation of gametes using laser microbeams. , 1991, Human reproduction.

[25]  C. Versaci,et al.  Andrology: Seventeen live births after the use of an erbium-yytrium aluminium garnet laser in the treatment of male factor infertility , 1994 .

[26]  R. Edwards,et al.  Maturation in vitro of human ovarian oöcytes. , 1965, Lancet.

[27]  W Feichtinger,et al.  Successful clinical application of laser for micromanipulation in an in vitro fertilization program. , 1992, Fertility and sterility.

[28]  Birth of a normal girl after in vitro fertilization and preimplantation diagnostic testing for cystic fibrosis. , 1992 .

[29]  A. H. Handyside,et al.  Pregnancies from biopsied human preimplantation embryos sexed by Y-specific DNA amplification , 1990, Nature.

[30]  Peter Fuhrberg,et al.  Photoablation of oocyte zona pellucida by erbium-yag laser for in-vitro fertilisation in severe male infertility , 1992, The Lancet.

[31]  K Rink,et al.  Microdissection of mouse and human zona pellucida using a 1.48-microns diode laser beam: efficacy and safety of the procedure. , 1995, Fertility and sterility.

[32]  J. Harper Preimplantation diagnosis of inherited disease by embryo biopsy: An update of the world figures , 1996, Journal of Assisted Reproduction and Genetics.