Human Ova Research Articles

Infection control considerations for in vitro fertilization and embryo transfer programs.

In vitro fertilization and embryo transfer (IVF-ET) is a process in which human ova are obtained by laparoscopic ovarian follicular puncture, fertilized in vitro by capacitated donor sperm, and introduced transcervically into the uterus. The prospective mother's ovarian cycle is usually stimulated with either clomiphene citrate, human menopausal gonadotropin, or both, so that multiple aspiratable follicles are produced. Follicular development is monitored by ultrasound and by serial serum or urine estrogen determinations. When adequate follicular development has been achieved, a pre-ovulatory dose of human chorionic gonadotropin is administered.

Fertilization of the Human Egg and Growth of the Human Zygote in vitro: The Singapore Experience

Human ova were collected by laparoscopy and matured in T6 medium supplemented with 10% heat‐inactivated serum for 4–6 hours. The semen was collected as split ejaculates at about the same time as the ova recovery and the better sample was used. The semen was prepared such that spermatozoa were obtained at room‐temperature, and a further sample obtained from the pellet at 37°C. One‐tenth to one‐half of a million 37°C layered spermatozoa were used for insemination of each egg. Sixteen to twenty hours later, the surrounding corona were broken down with either fine pipetting or needles, and the pronuclei identified. They were examined for cleavage sixteen to twenty‐four hours later. Replacement of the embryos, if fertilised and cleaved, was done within forty‐eight to seventy‐two hours of the ova recovery. The Bourn Hall catheter was used for the replacement, with the embryos suspended in 75% heat‐treated serum with T6. Of a total of 46 ova aspirated from patients with tubal obstruction, 34(73.9%) fertilized and divided. For patients with associated oligospermia, of 14 ova aspirated 9 (64.3%) fertilized and cleaved. For patients with prolonged idiopathic subfertility, of 22 ova aspirated 8 (36.4%) fertilized and cleaved. For patients with endometriosis, of 4 ova aspirated 3 (75%) fertilized and cleaved.

Effect of human follicular fluid on the maturity of rabbit ova by in vitro conditions

The influence of human follicular fluid from mature and immature oocytes on the maturation of rabbit oocytes under in-vitro conditions was examined. The human follicular fluids were obtained in the course of laparoscopies for extracorporeal insemination and separated into the categories mature and immature ova. In cases of normal early embryonal development the follicular fluid was termed "follicular fluid of a mature ovum"; if the ova were immature or degenerated the fluid was termed "follicular fluid of an immature ovum". Rabbit oocytes were obtained 3 hours after administration of P-LH and immediately cultivated in 3 different culture media for 7 hours. Only 6 out of 50 rabbit oocytes cultivated in Brackett's Defined Medium with the addition of 20% follicular fluid from an immature human ovum for 7 hours reached prometaphase II to metaphase II. This corresponds to a rate of 12%; on the other hand, with the addition of follicular fluid from a mature ovum 18 out of 50 oocytes reached the same phase, i.e., a rate of 36%. This difference is statistically significant (p less than 0.01). Brackett's Defined Medium with no fluid added was used as a control. Only 5 out of 50 oocytes managed to reach prometaphase II to metaphase II. The rate in this case was 10%. In order to prove that the rabbit oocytes were in the chromatin consolidation phase, they were fixed immediately after being obtained (3 hours after P-LH administration) and stained in 5% Giemsa solution. All of the oocytes (31) were in the chromatin consolidation phase.(ABSTRACT TRUNCATED AT 250 WORDS)

Fentanyl Does Not Inhibit Fertilization or Early Development of Sea Urchin Eggs

Sea urchin (Lytechinus variegatus) eggs have been used to study the effects of fentanyl on in vitro fertilization and early development as a model of human in vitro fertilization. Fentanyl did not affect fertilization or subsequent cell division when present in concentrations calculated to approximate or exceed those to which human ora would be exposed during clinical anesthesia. Lytechinus eggs exposed to fentanyl, then washed in fentanyl-free sea water before fertilization were also not affected and divided normally. The results suggest potential clinical utility of fentanyl during the harvesting of human ova for subsequent in vitro fertilization.

John Rock, developer of contraceptive pill, dies

John Rock, MD, the first scientist to perform in vitro fertilization of a human ovum in a test tube and one of the developers of the oral contraceptive, has died of a heart attack at the age of 94 years. He was a devout Roman Catholic, married in 1925 to Anna Thorndike by William Cardinal O'Connell of Boston. Rock subsequently became the father of four daughters and a son, and grandfather of 19. In the early 1920s, Rock began practicing obstetrics and gynecology in Boston and in 1924 he started the Fertility and Endocrine Clinic—one of the nation's first—at the Free Hospital for Women in Brookline, Mass. In 1944, he and Mirian F. Menkin, MD, conducted the first fertilization of a human ovum outside the body. Rock was best known for his work with Gregory Pincus, MD, and M. C. Chang, MD, in producing and clinically testing the oral contraceptive,

Legal Issues in Nonsurgical Human Ovum Transfer

Legal Issues in Nonsurgical Human Ovum Transfer

Legal issues in nonsurgical human ovum transfer.

Legal issues in nonsurgical human ovum transfer.

Zona-free hamster ovum penetration assay as a screening procedure for in vitro fertilization

Twenty couples with primary sterility due to mechanical factors entered our program of IVF. In addition to routine sperm evaluation, an SPA was performed in all cases. Whenever the SPA was pathologic, none of the human ova were fertilized in vitro, while 77% of the patients with a normal SPA had IVF of at least one human ovum. The good correlation between pathologic SPA and failure of IVF stresses the clinical value of this assay. The use of the SPA for patient selection for IVF is suggested.

Human ovum recovery via operative laparoscopy and in vitro fertilization

Between January 1979 and December 1980,142 laparoscopies were performed on 114 patients with long-standing tubal factor infertility. Twenty-eight were performed during spontaneous menstrual cycles, 20 following continuous human menopausal gonadotropin/human chorionic gonadotropin (hMG/hCG) therapy, and 94 following intermittent hMG/hCG stimulation. Follicular ripeness was judged by multiple criteria, which were also used to time the laparoscopy. Operative procedures were performed in all cases and follicular puncture was attempted in all but 16 subjects, where adhesions prevented access to the ovaries. In 28 spontaneous cycles, 28 follicles were punctured and 17 ova recovered; whereas in the 98 patients where ovulation was stimulated, 217 follicles were punctured and 43 ova collected. Following in vitro fertilization with the husband’s spermatozoa and embryo culture, pronucleus formation, 2-cell, 4-cell, 8-cell, and 16-cell stages were observed in 18 oocytes obtained from 17 patients. Embryo transfer has, however, not yet been performed.

Human ovum recovery via operative laparoscopy and in vitro fertilization

Between January 1979 and December 1980, 142 laparoscopies were performed on 114 patients with long-standing tubal factor infertility. Twenty-eight were performed during spontaneous menstrual cycles, 20 following continuous human menopausal gonadotropin/human chorionic gonadotropin (hMG/hCG) therapy, and 94 following intermittent hMG/hCG stimulation. Follicular ripeness was judged by multiple criteria, which were also used to time the laparoscopy. Operative procedures were performed in all cases and follicular puncture was attempted in all but 16 subjects, where adhesions prevented access to the ovaries. In 28 spontaneous cycles, 28 follicles were punctured and 17 ova recovered; whereas in the 98 patients where ovulation was stimulated, 217 follicles were punctured and 43 ova collected. Following in vitro fertilization with the husband's spermatozoa and embryo culture, pronucleus formation, 2-cell, 4-cell, 8-cell, and 16-cell stages were observed in 18 oocytes obtained from 17 patients. Embryo transfer has, however, not yet been performed.

Ultrastructural evaluation of 8–16 cell human embryos cultured in vitro

The fine structure of eight human embryos cleaving in culture are described. Preovulatory human oocytes aspirated at diagnostic laparoscopies were fertilized and developed in vitro by methods which produced normal pregnancies. Three 8-cell and five 10–16 cell embryos showing delayed cleavage were fixed, serially sectioned and examined to determine whether the embryos were developing normally or otherwise. The organization of two 8-cell embryos was apparently normal while the other embryos showed varying aspects of abnormal development. Most blastomeres contained organelles normally present in human ova and their fine structure closely resembled those of comparable mammalian embryos. Nearly all the cellular components encountered in early mammalian embryos were observed. Certain morphogenetic changes were also noted during early cleavage involving nuclei, smooth endoplasmic reticulum and mitochondria. Both normal and abnormal features of the embryos are reported. Multi-nucleated blastomeres and partial fragmentation were commonly seen in abnormal embryos. The importance of ultrastructural evaluation of embryos in an in vitro programme is revealed in this investigation.

Diploid nuclear replacement in mature human ova with cleavage

Diploid nuclear replacement in mature human ova with cleavage

Progress in Infertility

In the 1968 edition of this textbook, it was the intention of the editors to emphasize specific research aspects of infertility in the fields of genetics, reproductive biology, immunology, endocrinology, and surgery. The second edition contains 49 chapters with 60 contributing authors and continues to pursue that intention, making several noteworthy changes. The sections Ovarian Factors and Ovulation contain three new chapters. The first deals with the diagnosis and therapy of gonadal abnormalities. This frequently confusing topic is discussed in a clear and concise manner. The second section discusses human embryo transfer. It is exceptionally well written and is an excellent review of current information dealing with this controversial subject. The third, Implantation of the Human Ovum, is well illustrated, and the material is covered succinctly and thoroughly. In the section Endocrine Relationships, the chapters involving assay methods for pituitary, steroid, and thyroid hormones have been rewritten or significantly revised,

Example of a human ovum in a cervicovaginal smear fact or artefact?

A further example in a cervicovaginal smear of a structure with the morphological features of an unfertilized human ovum is reported. Previously published case reports of similar objects are reviewed, and alternative sources of bodies which could be incorrectly identified as ova are discussed together with suggested methods of establishing precise identity.

Ultrastructural studies of the implantation process.

The human ovum arrives at the uterine cavity by day 4 of development. At that time the ovum is a morula or an early blastocyst. Hertig et al. (1954) have described a human morula of about 96 h developmental age found within the uterine cavity and consisting of 58 cells: 5 embryonic and 53 trophoblastic. All the trophoblastic cells were of a fairly uniform cuboidal nature. The zona pellucida was still present (Carnegie No. 8794). In Hertig’s paper is a description of a human blastocyst of 4½ days of developmental age and consisting of 107 cells: 8 embryonic and 99 trophoblastic. An embryonic disc was formed at this stage. The trophoblastic cells were thinned out, except those placed at the embryonic pole. These cells (the “polar trophoblasts”) make the first contact with the uterine epithelium at the implantation site.

Fertilization of human ova in vitro.

Fertilization of human ova in vitro.

THE SEA URCHIN EGG AS A TESTING OBJECT IN TOXICOLOGY

Abstract: The sea urchin egg offers unique properties for experimental work on the cellular and embryologic level. The developmental stages are well suited for the study of teratogenic effects exerted by drugs. The development of the sea urchin egg is holoblastic and resembles the early development of the human ovum considerably. The gametes and embryos of the regular sea urchin species Paracentrotus lividus and Psammechinus microtuberculatus have been used in a study of the teratogenic effects of chloramphenicol, nicotine, chlorpromazine, imipramine and thalidomide. All the substances tested were found to influence the fertilization, cleavage and differentiation of the larvae. The effects of the substances on the cellular and embryonic level are described and the possible mechanism of action is discussed.During the past decade the interest in substances inflicting malformations or general injuries in living organisms has gained an increasing interest. The extensive use of biocides and their repercussions on living organisms of immediate economic interest and on man created an extending research in this field. Of particular importance was, however, the discovery of the teratogenic properties of the sedative hypnotic drug thalidomide. In spite of the elaborate pharmacological methods applied in testing drugs before their adoption in clinical use thalidomide, if administered during pregnancy, was ascertained to cause congenital malformations in man. This was a surprise since thalidomide was regarded as a particularly safe drug and even the intake of large overdoses did not seem to cause any persisting ill effects. There is, however, some confusion about the teratogenic effect of thalidomide and its mode of action is still obscure.In the present investigation a number of substances with known or suspected teratogenic effect in man were studied on the cellular and embryonic level. As material served the gametes and larvae from the regular sea urchin Paracentrotus lividus (LAMARCK), which is the classic material for the study of fertilization, cleavage and development. Few organisms have been subjected to a more conclusive experimental work, and the main advantage with this material is that every ripe female delivers several millions of fairly uniform transparent eggs, particularly suited for microscopic examination in the living state. Subsequent to insemination the eggs develop within the course of a few days into self‐maintaining larvae. Cleavage and organogenesis are also easy to observe and even minor changes taking place after the addition of an active drug can be registered in the microscope.The fact that the cleavage and embryogenesis are so well investigated also makes it possible to study the specific effect on e.g. the ectoderm or the endoderm of a certain substance. The short period of time necessary for the embryo to reach the self‐maintaining pluteus stage also minimizes the influence of external experimental factors.

Ultrastructure of a Human Implantation Site

The fins structure of the trophoblast from a previllous human ovum is described. The extra‐embryonal mesoderm was lined by cylotrophoblasts, which were beginning to form the primary villi. The ultrastructure of these cells was similar to that of the Langhans' cells of the definitive villus. The syncytiotrophoblast was located peripherally and consisted of a mixture of giant cells and syncytial masses. This cell type was well differentiated, with its cytoplasm occupied by large amounts of granular endoplasmic reticulum. Golgi elements, and vesicles. The syncytiotrophoblast had many microvilli and pinocytotic vesicles at the surface, lining the lacunae which contained maternal blood cells. Close contact was observed between the trophoblast and the cells of the maternal glands. The trophoblastic invasion of a maternal vessel is described, and the trophoblastic invasion is discussed.

Cleavage of Human Ova in Vitro

Cleavage of Human Ova in Vitro

Observations on human ova and the fertilization process

The mammalian fertilization process was reviewed, with emphasis on recent laboratory developments. Maturation stages of the human follicular oocyte which are requisite to fertilization were explored with in vitro culture methods. The experience of other investigators was confirmed in that, on removal from the follicle, nuclear maturation through polar body release was observed. Available information on the human tubal ovum was summarized and the recorded attempts at in vitro fertilization of the human oocyte analyzed. Time lapse in cinematography used successfully in the rabbit was suggested as a means for establishing the validity of in vitro fertilization in man. It was predicted, based on experience in other mammals, that placement of in vitro cultured human oocytes into the uterus for implantation would eventually be carried out successfully. Further studies in the subhuman primate were suggested as a requisite to such experiments, in order to insure conditions which would yield normal young.