abberior dyes & labels
2024
Journal of Assisted Reproduction and Genetics
Rescue in vitro maturation using ovarian support cells of human oocytes from conventional stimulation cycles yields oocytes with improved nuclear maturation and transcriptomic resemblance to in vivo matured oocytes
Authors:
Bruna Paulsen, Sabrina Piechota, Ferran Barrachina, Alexa Giovannini, Simone Kats, Kathryn S. Potts, Graham Rockwell, Maria Marchante, Samantha L. Estevez, Alexander D. Noblett, Alexandra B. Figueroa, Caroline Aschenberger, Dawn A. Kelk, Marcy Forti, Shelby Marcinyshyn, Klaus Wiemer, Marta Sanchez, Pedro Belchin, Joseph A. Lee, Erkan Buyuk, Rick E. Slifkin, Merrick Pierson Smela, Patrick R. J. Fortuna, Pranam Chatterjee, David H. McCulloh, Alan B. Copperman, Daniel Ordonez-Perez, Joshua U. Klein & Christian C. Kramme
Keywords:
Ovarian support cells; In vitro maturation; Stem cells; Oocyte transcriptomics; Granulosa cells
Abstract:
Purpose Determine if the gene expression profiles of ovarian support cells (OSCs) and cumulus-free oocytes are bidirectionally influenced by co-culture during in vitro maturation (IVM).
Methods Fertility patients aged 25 to 45 years old undergoing conventional ovarian stimulation donated denuded immature oocytes for research. Oocytes were randomly allocated to either OSC-IVM culture (intervention) or Media-IVM culture (control) for 24–28 h. The OSC-IVM culture condition was composed of 100,000 OSCs in suspension culture with human chorionic gonadotropin (hCG), recombinant follicle stimulating hormone (rFSH), androstenedione, and doxycycline supplementation. The Media-IVM control lacked OSCs and contained the same supplementation. A limited set of in vivo matured MII oocytes were donated for comparative evaluation. Endpoints consisted of MII formation rate, morphological and spindle quality assessment, and gene expression analysis compared to in vitro and in vivo controls.
Results OSC-IVM resulted in a statistically significant improvement in MII formation rate compared to the Media-IVM control, with no apparent effect on morphology or spindle assembly. OSC-IVM MII oocytes displayed a closer transcriptomic maturity signature to IVF-MII controls than Media-IVM control MII oocytes. The gene expression profile of OSCs was modulated in the presence of oocytes, displaying culture- and time-dependent differential gene expression during IVM.
Conclusion The OSC-IVM platform is a novel tool for rescue maturation of human oocytes, yielding oocytes with improved nuclear maturation and a closer transcriptomic resemblance to in vivo matured oocytes, indicating a potential enhancement in oocyte cytoplasmic maturation. These improvements on oocyte quality after OSC-IVM are possibly occurring through bidirectional crosstalk of cumulus-free oocytes and ovarian support cells.