Use of fluorescence in situ hybridization to assess the chromosomal status of embryos obtained from cryopreserved oocytes

Abstract

Objective: To analyze the chromosomal status of human embryos obtained from frozen-thawed oocytes. Design: Fluorescence in situ hybridization analysis of embryos obtained after oocyte cryopreservation. Setting: Department of Obstetrics and Gynecology at the University of Perugia, Italy, and the Instituto Valenciano de Infertilidad, Spain. Patient(s): Oocyte donors ( n = 43). Fertilization, development, and chromosomal status of the embryos were compared with a control group ( n = 18) of patients undergoing preimplantation genetic diagnosis for sex chromosome–linked diseases. Intervention(s): Collection of oocytes after conventional ovarian stimulation and cryopreservation using propanediol as the cryoprotectant and a slow freezing procedure. Microinjection of surviving metaphase II oocytes and evaluation of fertilization and embryo development up to blastocyst stage. Chromosomal analysis after embryo biopsy. Main Outcome Measure(s): Survival, fertilization, and blastocyst rates. Embryo chromosomal analysis employing specific probes for chromosomes 13,18,21, X and Y. Result(s): The overall survival rate was 59.4%. There was no difference between cryopreservation and control groups in fertilization rates (76.5% vs. 90.5%) or blastocyst development (29.6% vs. 35%). The percentage of blastocysts from the original number of cryopreserved oocytes was only 5.6%, comparable to the 5.9% obtained in the control group. The percentage of embryos with abnormal number of chromosomes in the cryopreservation group (28.6%) was comparable to the 26% observed in the controls. Conclusion(s): Fertilization and cleavage rates after oocyte freezing are acceptable. Survival is, however, still poor, leading to overall results that make the technique clinically inefficient. There is no increase in the rate of chromosomal abnormalities, indicating that the technique is, nevertheless, safe enough to be further explored and improved.