Martin M. Quigley

Hormonal regulation of human endometrial stromal cells in culture: an in vitro model for decidualization

Stromal cells derived from proliferative or secretory human endometria, cultured in the absence of steroid hormones, grew as monolayers that showed only occasional areas of immunoreactive fibronectin and did not produce detectable levels of prolactin (PRL) or laminin. Treatment with physiological doses of estradiol and progesterone induced PRL production and stimulated cell proliferation, resulting in multilayering with an increase of the saturation density. Electron microscopy showed the development of gap junctions, whereas immunofluorescence revealed a dense pericellular matrix containing fibronectin and laminin. These findings show that human endometrial stromal cells in culture respond to physiological doses of ovarian hormones with ultrastructural, proliferative, and biochemical changes that are characteristic of decidualization in vivo. This culture system thus provides an in vitro model for human decidualization.

Laboratory setup for human in vitro fertilization

None of the procedures utilized in human IVF are unique or technically very difficult to perform. However, when considering the nature of the material being handled, it is essential that all laboratory procedures, equipment, and supplies are optimal for IVF . The individuals handling the gametes and embryos must be suitably trained and experienced in handling mammalian gametes and embryos before working with human tissues. In addition, the technologists must be trained in proper tissue culture techniques. Above and beyond the training and experience of the technologist, it is necessary that all laboratory equipment and chemicals for medium preparation be of the highest quality and known consistency, and that periodic checks of the culture conditions be performed using the bioassay of mouse embryos. The functioning of the equipment must be constantly monitored and backup systems be in place wherever possible in order to minimize the chances that laboratory accidents will result in loss or damage of the embryos. Meticulous attention to all aspects of the laboratory procedures will result in the patients having the best possible chance to obtain a pregnancy.

Follicular aspiration: a comparison of an ultrasonic endovaginal transducer with fixed needle guide and other retrieval methods

Ultrasonic techniques have been developed as alternatives to conventional laparoscopic aspiration for oocyte retrieval for in vitro fertilization (IVF). Given the advantages (less risk, lower cost, and greater patient acceptance) of these alternative techniques, it is appropriate to assess their efficacy compared with traditional laparoscopic retrieval. This article examines the recovery rate of oocytes and their subsequent fertilization rate with the use of an ultrasonic endovaginal transducer with fixed needle guide and compares these results with other retrieval methods. Comparisons were made between laparoscopic harvesting (n = 71, group I), ultrasonic transabdominal transvesical (n = 21, group II), and ultrasonic vaginal transducer (n = 76, group III). The data demonstrate comparable success using an ultrasonic endovaginal transducer with fixed needle guide. The authors believe this technique to be the procedure of choice for all routine oocyte retrievals during IVF treatment.

Human in vitro fertilization and embryo transfer

1 Historical Background and Essentials for a Program in In Vitro Fertilization and Embryo Transfer.- 2 Legal Issues Raised by In Vitro Fertilization and Embryo Transfer.- 3 Patient Screening and Selection.- 4 Hypothalamic Control of the Menstrual Cycle.- 5 Maturation of the Follicular Microenvironment.- 6 Enhanced Follicular Development with Clomiphene Citrate and Human Chorionic Gonadotropin.- 7 Human Menopausal Gonadotropins for Follicular Recruitment in In Vitro Fertilization and Embryo Transfer.- 8 Monitoring Follicular Development with Ultrasound.- 9 Monitoring Follicular Development with Estrogens.- 10 Laparoscopic Follicular Aspiration.- 11 Sperm Capacitation.- 12 Oogenesis, Fertilization and Early Development.- 13 Laboratory Details in an In Vitro Fertilization and Embryo Transfer Program.- 14 Fertility Potential Evaluation with the Zona-free Hamster Egg Bioassay.- 15 Techniques of Embryo Transfer.- 16 Implantation.- 17 Pregnancy Management Following In Vitro Fertilization and Embryo Transfer.- 18 The Clinical Coordinator in an In Vitro Fertilization and Embryo Transfer Program.- 19 Data Management in an In Vitro Fertilization and Embryo Transfer Program.- 20 Ethical Considerations in In Vitro Fertilization and Embryo Transfer.- Appendices.- Contributors.

A program for matched, anonymous oocyte donation.

The authors program for matched, anonymous oocyte donation has resulted in two successful pregnancies among the first eight oocyte recipients. All oocyte recipients to date have had ovarian failure or absence with premature ovarian failure the most common cause. All recipients were cycled on a program of incremental oral micronized estradiol and intramuscular progesterone-in-oil. Thirteen candidates for oocyte donation were screened to obtain 8 donors. One donor candidate was excluded because of her medical history. The psychological screening of 2 of the other donor candidates (who subsequently did not complete the donation cycle) revealed a primary motive of financial gain. In general, the psychological profiles of donor candidates revealed a high incidence of troubled families and either reproductive loss or loss of a parent. Ovarian stimulation of the donors followed our standard in vitro fertilization protocol. The recipients exogenous steroid replacement continued until days 97 and 101, respectively, of the two gestations. Both pregnancies resulted in the delivery of normal singleton males--the first at 40 weeks, the second at 35 weeks.

Estrogen replacement therapy: help or hazard?

FEW controversies in the practice of medicine have generated as much discussion as has the role of estrogen-replacement therapy in the climacteric woman. The literature is replete with articles abo...

Timing human chorionic gonadotropin administration by days of estradiol rise

A critical aspect of human in vitro fertilization (IVF) treatment programs is the timing of human chorionic gonadotropin (hCG) administration to induce final oocyte maturation. Patients were treated with clomiphene citrate (Serophene, Serono Laboratories, Inc., Randolph, MA), 100 mg, and human menopausal gonadotropin (Pergonal, Serono Laboratories, Inc.) 1 ampule daily from cycle days 4 through 8. The baseline estradiol (E2) level was determined by averaging the E2 values from cycle days 3 and 4. The first day of E2 rise was defined as the day that the E2 level exceeded 150% of the baseline. In general, 5000 IU of hCG (Profasi-HP, Serono Laboratories, Inc.) was administered on the evening of the sixth day of E2 rise. Significantly fewer oocytes were recovered per patient when the hCG was administered on the seventh day of E2 rise, compared with the fifth or sixth day. The recovered oocytes in the seventh-day E2 group had a significantly lower fertilization rate, compared with the other groups. The use of E2-rise days proved to be a simple, successful technique for the timing of hCG administration in an IVF treatment program.

Preliminary experince with a combination of clomiphene and variable dosages of menopausal gonadotropins for enhanced follicular recruitment

A combination of clomiphene citrate and human menopausal gonadotropin was employed for enhanced follicular recruitment in an in vitro fertilization program. All patients received 50 mg of clomiphene and 1 ampule of human menopausal gonadotropin daily from cycle day 5 through cycle day 9. Follicular monitoring was begun on day 10 using a combination of ultrasound measurement of follicular size and number and determination of peripheral estradiol levels. Based on the size and number of follicles, the peirpheral levels of estradiol, and the rate of follicular growth and increase in estradiol, human menopausal gonadotropin was continued at a dosage of 1 to 3 ampules/day through the day of human chorionic gonadotropin administration. Human chorionic gonadotropin was administered on the evening of the day the largest follicle reached or exceeded 20 mm in mean diameter if the estradiol levels had been rapidly rising or reaching a plateau and had exceeded a minimal level of 300 pg/ml. Using this protocol, 30 of 33 patients underwent laparoscopy, 29 patients had successful oocyte recovery, and 23 patients underwent embryo replacement, with the establishment of six clinical pregnancies.

Simultaneous in vitro fertilization and gamete intrafallopian tranfer (GIFT)

During a 6-month period, a combination of gamete intrafallopian transfer (GIFT) and in vitro fertilization (IVF) was offered to all couples beginning an IVF treatment cycle in whom the wife had anatomically normal fallopian tubes. It was recommended to these couples that sufficient oocytes be reserved for insemination in vitro to determine whether the husbands spermatozoa could fertilize the wifes oocytes. During this interval, 16 couples underwent the combined IVF-GIFT procedure. All of the IVF-GIFT couples had at least two oocytes inseminated in vitro and at least two oocytes for GIFT. Of the 16 IVF-GIFT couples, only 1 (6.25%) achieved a clinical pregnancy. More important, 50% (8/16) of the IVF-GIFT couples had no oocytes fertilized in vitro. With the information concerning lack of fertilization in vitro, appropriate recommendations concerning future fertility management can be made. If the same couples had undergone the GIFT procedure alone, without additional oocytes fertilized in vitro, this information would not have been obtained.

Human in vitro fertilization and the law

Physicians who intend to perform in vitro fertilization--or the newer alternative, in vivo fertilization and embryo transfer--should be aware of the plethora of laws that potentially cover their work. In perhaps no other area of medicine are there so many separate statutes and regulations that potentially apply. State fetal research laws, abortion statutes, human subject protection laws, and specific in vitro fertilization statutes can determine whether and how the procedure can be undertaken. When donor gametes or a surrogate carrier is used, additional laws governing artificial insemination, paternity, or adoption may come into play to determine the childs legal status and its relationship to the parties involved. This article is designed to guide physicians through the legal maze.