Production of embryos from in vitro-matured primary human oocytes.

OBJECTIVE: To determine the factors that influence the number and quality of embryos produced from primary oocytes collected from untreated regularly ovulating and irregular or anovulatory polycystic women. DESIGN: A direct comparison between two patient groups whose oocytes were matured in vitro and a comparison of the embryo development of in vitro-matured oocytes from untreated patients with in vivo-matured oocytes of superovulated IVF-ET patients obtained during the same period. SETTING: The Monash IVF Clinic, involving patients who expressed the desire to avoid super-ovulation with fertility drugs. MAIN OUTCOME MEASURES: The completion of nuclear maturation of oocytes after 36 or 48 hours culture, fertilization in vitro, and embryo development ratio. RESULTS: Oocytes from regular cycling patients matured and fertilized at significantly higher rates than irregular cycling and anovulatory women and their embryos had significantly higher mean embryo development ratio. The mean embryo development ratio of embryos of regular cycling patients was similar to superovulated IVF patients but irregular cycling and anovulatory patients had a significantly lower embryo development ratio. Culture of oocytes for 48 hours increased maturation of oocytes from 57% to 82% but did not affect fertilization or cleavage rates. Embryo development was not affected significantly by the grade of follicular cell cover of oocytes. CONCLUSIONS: The developmental capability of primary oocytes is higher in regular cycling women than in irregular cycling and anovulatory women with polycystic ovary disease.

Blastocyst development and birth after in-vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching.

Immature oocyte recovery followed by in-vitro oocyte maturation and in-vitro fertilization is a promising new technology for the treatment of human infertility. The technology is attractive to potential oocyte donors and infertile couples because of its reduced treatment intervention. Immature oocytes were recovered by ultrasound-guided transvaginal follicular aspiration. Oocytes were matured in vitro for 36-48 h followed by intracytoplasmic sperm injection (ICSI). Embryos were cultured in vitro for 3 or 5 days before replacement. Assisted hatching was performed on a day 5 blastocyst stage embryo. Embryo and uterine synchrony were potentially enhanced by luteinization of the dominant follicle at the time of immature oocyte recovery. Mature oocyte and embryo production from immature oocyte recovery were similar to the previous IVF results of the patients. A blastocyst stage embryo, produced as a result of in-vitro maturation, ICSI, in-vitro culture and assisted hatching, resulted in the birth of a healthy baby girl at 39 weeks of gestation.

Determination of the charge content at the surface of cells using a colloid titration technique.

A colloid titration technique has been used to estimate the surface charge content of three distinct cell types of differing surface charge characteristics, i.e., human red blood cells, the surface of which is studded with sialic acid residues, endothelial cells which are surrounded by a thick glycocalyx, and chondrocytes which, when grown at high cell density for several weeks, synthesize a dense pericellular matrix similar to that observed in cartilaginous tissues. Estimates of the charge content obtained for human erythrocyte ghosts and cultured endothelial cells are in good agreement with the charge determined by chemical analyses. On the other hand, the charge at the surface of chondrocytes represented only a fraction of that calculated from measurements of the glycosaminoglycan content (15% by Day 12 in culture); close correlation between charge and chemical amount could be obtained only after digestion of the cell layer with collagenase and papain indicating the possible electrostatic involvement of the negative groups of the glycosaminoglycan chains with basic proteins. Thus, the colloid titration technique may provide a simple and sensitive tool to study the interactions occurring between the extracellular matrix components while the matrix is being formed and to establish the chemical nature of the molecules contributing to the cell surface charge.