In vitro maturation of human oocytes in a follicle-mimicking three-dimensional coculture

Objective: To verify the hypothesis that a three-dimensional, follicle-mimicking structure enhances in vitro maturation yields without hormonal supplementation in an in vitro maturation program. Design: Feasibility study; 204 anonymous denuded germinal vesicles retrieved from gonadotropin-treated women were cultured for 48 hours without hormonal supplementation in microdrop culture or in a three-dimensional coculture with granulosa cells in a barium alginate membrane. Setting: An assisted reproduction center in Italy. Patient(s): One hundred twenty-two informed women. Intervention(s): Germinal vesicles retrieved after ovarian stimulation were denuded and cultured without hormonal supplementation in microdrop culture or in a three-dimensional coculture with granulosa cells enclosed in the core of a barium alginate capsule. Main Outcome Measure(s): Oocyte maturation and morphological investigation of follicle-mimicking structures. Result(s): Statistically significantly higher oocyte maturation yields were obtained by using the three-dimensional coculture system enclosed in a barium alginate membrane (after 48 hours: coculture, 90.3%; microdrop, 52.0%; odds ratio, 8.51). Morphological investigation indicated that after 48 hours of coculture, the decumulated oocyte appeared to be surrounded by a pseudocumulus structure that lies on the inner surface of the alginate membrane and protrudes toward the capsule's lumen. Conclusion(s): The maturation yield of cocultured oocytes is eightfold higher than that achieved with a microdrop maturation technique, making a higher number of gametes available for IVF programs. Stereomicroscopic morphological survey indicates a reassembly of granulosa cells surrounding oocyte zona, mimicking an antral cumulus oophorus.