Uses and limitations of two molecular cytogenetic techniques for the study of arrested embryos obtained through assisted reproduction technology

We studied chromosomal abnormalities in arrested embryos produced by assisted reproductive technology with fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH) in order to determine the best technique for evaluating chromosomal aneusomies to be implemented in different situations. We examined individual blastomeres from arrested embryos by FISH and arrested whole embryos by CGH. All of the 10 FISH-analyzed embryos gave results, while only 7 of the 30 embryos analyzed by CGH were usable. Fifteen of the 17 embryos were chromosomally abnormal. CGH provided more accurate data for arrested embryos; however, FISH is the technique of choice for screening in preimplantation genetic diagnosis, because the results can be obtained within a day, while the embryos are still in culture.

Birth resulting from transfer of blastocysts cryopreserved with propanediol after spontaneous hatching.

This paper reports the birth of a healthy baby resulting from transfer of blastocysts that were cryopreserved using propanediol after spontaneous hatching. A young infertile couple underwent IVF treatment in the clinic. After several IVF attempts, two births resulted; the first one with fresh embryos in 1996 after three IVF cycles, and the second one in 1999 (after a new IVF cycle in 1998) with frozen blastocysts that had remained cryopreserved in 1.5 mol/l propanediol and 0.1 mol/l sucrose after spontaneous hatching. This report of a healthy baby following transfer of hatched blastocysts frozen in propanediol supports further exploration of this approach.

Clinical practice of embryo transfer.

In this review, several embryo transfer methods are considered, together with factors involved in achieving an effective transfer. The approach most used is transcervical intrauterine transfer. This is described in detail, together with the many variables influencing success, e.g. technical ability and training of personnel, catheter choice, value of a previous 'dummy transfer' and the need to minimize trauma during transfer and so prevent damage to the uterine lining, bleeding and uterine contractions. These factors can each negatively impact on pregnancy rates. Emphasis is put on quality, developmental stage and number of embryos to be transferred to limit multiple pregnancies and their unwanted side-effects. Culture to blastocyst stages and single embryo transfer when optimal quality embryos are available are discussed as means of avoiding multiple pregnancies. Reference is made to embryo cryopreservation and fertility following frozen embryo transfer. Other techniques, such as ultrasound-controlled transcervical intrauterine transfer, and ultrasound-controlled transmyometrial transfer, are reviewed. More invasive procedures, generically grouped as surgical embryo transfer, including gamete intra-Fallopian transfer (GIFT), zygote intra-Fallopian transfer (ZIFT), pronuclear stage transfer and embryo intra-Fallopian transfer (EIFT), are also described. These techniques had a place in IVF when the need to apply assisted reproductive techniques exceeded the capacity of most laboratories, but not today thanks to refined laboratory technology and improved understanding of implantation. Alternative assisted reproductive technologies, such as direct intra-follicular insemination (DIFI), Fallopian spermatic perfusion (FSP), peritoneal oocyte stage and sperm transfer and intra-vaginal culture (IVC), are mentioned briefly.

Empty follicle syndrome due to human errors: its occurrence in an in-vitro fertilization programme.

We report five cases in which no oocytes were retrieved after standard ovarian stimulation for in-vitro fertilization (IVF), and in which it was found that mistakes had been made at the time of human chorionic gonadotrophin (HCG) administration. In all five cases, oocyte retrieval was achieved after injecting HCG, when necessary, and reprogramming aspiration 24-36 h later. A mean of 7+/-3.2 MII oocytes were recovered per patient and 3.2+/-0.8 embryos were transferred. Three clinical pregnancies were obtained, and four healthy infants were born. In our programme, these were the only cases of empty follicle syndrome (EFS) that appeared over a total of 1118 cycles, and were all explained by human error in the administration of HCG. Our experience shows that human error could be considered a significant factor in the aetiology of empty follicle syndrome, and that EFS may be in part avoided by taking simple preventive measures.

[Pregnancy obtained by intracytoplasmic oocyte injection of spermatozoa retrieved from a cryopreserved testicular biopsy]. / Embarazo logrado por inyección intracitoplasmática de ovocitos con espermatozoides provenientes de una biopsia de testículo criopreservada.

We have obtained a clinical pregnancy in a 26 year old patient by means of in vitro oocyte fertilization by intracytoplasmic injection of spermatozoa obtained from a cryopreserved testicular biopsy. In a first attempt performed with fresh biopsy material, the woman became pregnant, but the pregnancy ended in a spontaneous abortion. In a second cycle, the spermatozoa were retrieved from a cryopreserved sample saved from the first attempt. Twelve metaphase II oocytes were collected and injected; from these, nine became fertilized, three preembryos were transferred at the eighth-cell stage and the other six were cryopreserved. An ongoing clinical pregnancy was obtained with two gestational sacs.