Andrew L. Speirs

The Ultrastructure of the Preovulatory Human Egg Fertilized In Vitro

Preovulatory eggs in cumulus were inseminated in vitro with washed spermatozoa which had been preincubated for 1.5 hours. After 3 hours, three eggs were processed for electron microscopy and each was sectioned serially from pole to pole. In the two eggs which had been fertilized, the expanded chromatin of the fertilizing sperm head and the chromatin of the ovum were almost completely surrounded by a developing pronuclear envelope. In one of the penetrated eggs the developing male pronucleus and associated midpiece and sperm tail were located within an incorporation cone. The surface of the cone was free of microvilli but contained a zone of microfilaments immediately beneath the plasma membrane. A similar zone of microfilaments was present beneath the advancing surface of the extruding second polar body (PB2) which was connected to the ovum by an interbody and microtubules of the meiotic spindle. Cortical granules were completely absent from the fertilized eggs but were present in the unfertilized egg. PB2 contained a nucleus at a stage of development similar to that of the early pronuclei.

Analysis of the benefits and risks of multiple embryo transfer.

Depending on embryo quality and recipient factors, increasing the number of embryos transferred after in vitro fertilization may produce more pregnant patients or more multiple pregnancies. An analysis is outlined demonstrating how observations on the placement of two embryos in the uterus may be used to estimate the risks and benefits associated with the transfer of multiple embryos. Currently, the highest pregnancy rates are obtained with multiple embryo transfers, and these results are compared with the mathematical model.

Serum supplement in human in vitro fertilization and embryo development

The effects of maternal preovulatory serum and human fetal cord serum supplement in culture medium in human in vitro fertilization and embryo development were compared in 208 cases over an 8-month period when there were no significant changes in other variables. A significantly higher pregnancy rate was observed in the fetal cord serum group despite no significant difference being found in fertilization and embryo cleavage rates. This suggests a difference in the health of the embryo cultured in different serum supplements.

Pregnancies after intracytoplasmic injection of sperm collected by fine needle biopsy of the testis

OBJECTIVE To investigate the use of intracytoplasmic sperm injection with sperm collected by fine needle biopsy of the testis as a treatment for male genital tract obstruction. DESIGN Sperm isolated from a fine needle biopsy of the testis were used to inseminate oocytes by intracytoplasmic sperm injection. SETTING A hospital-based tertiary referral infertility service. PATIENTS Case studies of two couples in whom the male partner had a genital tract obstruction. MAIN OUTCOME MEASURES Fertilization and pregnancy. RESULTS In the first case, 9 oocytes fertilized normally out of 13 injected. After the transfer of fresh (one cycle) and frozen (four cycles) embryos, a single intrauterine fetal heart pregnancy was achieved. In the second case, five oocytes fertilized normally from nine oocytes injected; two embryos were transferred fresh and three were frozen. A single fetal heart intrauterine gestation was obtained after the initial transfer of two fresh embryos. CONCLUSIONS A high normal fertilization rate and pregnancies are possible with intracytoplasmic sperm injection using sperm collected directly from the testis. Sperm retrieval by fine needle biopsy offers a viable alternative to microsurgical aspiration and is also suitable for treating patients with intratesticular blockage.

A laparoscopic approach to a program of gamete intrafallopian transfer

Gamete intrafallopian transfer (GIFT) provides an effective method of achieving pregnancy in infertile women with normal fallopian tubes. Laparoscopic approach to ovum pickup and tubal catheterization provides a simple and rapid means of performing the operation. Equipment used to facilitate this process is described, and techniques of tubal catheterization are discussed. A clinical pregnancy rate of 27% is reported in a series of 71 treatment cycles. The application of GIFT in conjunction with in vitro fertilization is discussed, especially the use of excess gametes to provide embryos for freezing. The use of GIFT as a research and investigative tool may provide further insight into the causes for idiopathic infertility.

Detection of Antispermatozoal Antibodies of IgA Class in Cervical Mucus

ABSTRACT: A simple procedure for detection of antisperm antibodies of IgA class in human cervical mucus is described and the results of its application to samples from 102 patients are presented. The results suggest that the IgA immunobead test (IgA‐IBT) is a specific and clinically useful test for sperm antibodies. There was a strong correlation between the IgA‐IBT and the presence of complement‐dependent sperm immobilization in serum (Spearmans, r = 0.92, p < 0.001). Positive IgA‐IBT results occurred only in mucus samples that showed poor penetration by normal sperm. An added advantage of the IgA‐IBT is that both the immunoglobulin class and the site of binding to the sperm surface can be determined simultaneously.

Performance of patients with a “frozen pelvis” in an in vitro fertilization program

It is now possible to identify and study the performance of different subgroups of patients in in vitro fertilization (IVF) programs. Patients with severe pelvic adhesions due to pelvic inflammatory disease (PID) or endometriosis were classed as having a frozen pelvis if less than or equal to 20% of total ovarian surface was visible and if the rest of the ovary was bound down with significant adhesions. IVF offers the only hope of pregnancy for these patients. Fifty-one treatment cycles in 23 such patients were matched against 51 cycles in 48 patients with adhesion-free ovaries. The study group had a significantly higher number of cancelled oocyte retrievals because of poor estradiol (E2) response. They also had a significantly lower rate of E2 rise and a lower peak value of E2 before and after the administration of human chorionic gonadotropin. These patients took longer to respond to a hyperstimulation regime, and when a response occurred they formed fewer follicles, as measured with the use of ultrasound. Lower numbers of oocytes were obtained from this group, but the fertilization rate of oocytes was the same for both groups. One pregnancy occurred in the study group and 11 in the control group. It is possible that disruption of ovarian blood supply or mechanical factors due to the pressure of significant adhesions prevent a good follicular response in patients with a frozen pelvis.

Ultrasound Control of Clomiphene/Human Chorionic Gonadotropin Stimulated Cycles for Oocyte Recovery and in Vitro Fertilization

Three pregnancies have been achieved through in vitro fertilization (IVF) following clomiphene/human chorionic gonadotropin (hCG) stimulation monitored only with ultrasound. When the results of 120 stimulated cycles were compared with those from 213 spontaneous cycles during a one-year period, the clomiphene-ultrasound-hCG method led to a significantly higher laparoscopy rate as well as significantly better yields of mature oocytes and embryos for intrauterine transfer. The luteal phase was normal in the stimulated group. This ultrasound-monitored technique was simpler to manage and less costly and appears to be the current method of choice for obtaining oocytes for IVF.

Increasing the human menopausal gonadotropin dose—does the response really improve?

This study assesses the effects of attempts to optimize human menopausal gonadotropin (hMG) dosage in 271 patients who had at least two hyperstimulation cycles for in vitro fertilization or gamete intrafallopian transfer. In the first cycle, all patients received clomiphene citrate and hMG 150 IU/d. In the second cycle, the hMG dose was increased in 45% of patients to try to increase the egg yield. In spite of the increase, the population response was practically identical in both cycles. Median numbers of eggs retrieved (6 versus 6), no eggs retrieved (0.4% versus 1%), only one or two eggs retrieved (10% versus 10%), and canceled cycles (10% versus 10.7%). This suggests that increasing the hMG dosage above 150 IU does not increase the number of eggs retrieved. A poor response may be due to inherent differences in follicular development that cannot be overcome by increases in hMG dosage.

Patient Selection for in Vitro Fertilization: Physical and Psychological Aspects

In vitro fertilization (IVF) and embryo replacement offers the infertile couple a whole new dimension in their treatment. It offers new hope to the previously abandoned, and it has sparked a veritable frenzy of scientific endeavor among the reproductive biologists of the world. It has attracted extensive media attention, and, as each technological breakthrough is announced, the euphoria increases and the horizons seem endless.