The protective effects of cord blood-derived exosomes on oocyte and ovarian granulosa cells during cisplatin chemotherapy

Islam M. Saadeldin^1*, Budur Alshehri¹, Maha AlThubiyani¹, Rana Alhamdan², Serdar Coskun², Ameera Gaafer³, Sheema Almozyan³, Zakia Shinwari⁴, Ayodele Alaiya⁴, Ayman Swelum5, Abdullah M. Assiri¹

¹Comparative Medicine Department, KFSH&RC (imohamed@kfshrc.edu.sa)

²IVF unit, Pathology and Laboratory Medicine Department, KFSH&RC

³Cell Therapy and Immunobiology Department, KFSH&RC

⁴Proteomics Unit, Stem Cell & Tissue Re-Engineering Program, KFSH&RC
5Department of Animal Production, College of Food and Agriculture Science, King Saud University


Abstract Text:

The study aims to evaluate the protective and ameliorative effects of umbilical cord blood-derived exosomes on oocytes and ovarian granulosa cells subjected to cisplatin chemotherapy. Using an ovine sheep model and human ovarian garnulosa cells, the research employed in vitro maturation (IVM) and intracytoplasmic sperm injection (ICSI) techniques to assess the therapeutic potential of exosomes. Moreover, human and sheep granulosa cells were used to assess the effects of cisplatin on cell growth and migration. Oocytes were cultured under four treatments: control, cisplatin, cisplatin with exosomes, and exosomes alone. Following maturation, ICSI was performed to measure fertilization and subsequent embryo development. Hoechst staining was used to identify successful fertilization (2PN stage), qPCR was used to study gene expression, and the TUNEL assay detected apoptosis in blastocysts.

Exosomes were characterized through transmission electron microscopy, nanoparticles tracking analysis, and proteomics for analyzing the protein cargo contents. The findings demonstrated that exosomes enhanced oocyte maturation, cumulus cells expansion, improved fertilization rates, and supported embryo development to the blastocyst stage, even under cisplatin-induced oxidative stress. Specifically, cisplatin-treated oocytes exhibited reduced cumulus expansion, nuclear maturation, and increased reactive oxygen species with disturbed gene expression while the addition of exosomes significantly mitigated these detrimental effects. Embryos from the cisplatin + exosome group showed improved cleavage and blastocyst formation compared to the cisplatin group. Furthermore, exosome-treated embryos displayed lower apoptosis and restored blastocyst total cell count, confirming their protective and restorative roles. Similarly, human and sheep granulosa cells treated with cisplatin + exosome showed increased ki-67 expression and increased wound healing capacity.

This study underscores the potential of cord blood stem cell-derived exosomes in reducing chemotherapy-induced ovarian damage by counteracting oxidative stress and promoting cellular recovery. The bioactive molecules within exosomes, such proteins, facilitate these reparative processes, paving the way for novel oncofertility strategies. The findings provide hope for preserving reproductive potential in cancer patients undergoing chemotherapy. These results highlight a promising foundation for innovative therapeutic approaches to fertility preservation, addressing a critical need in the field of reproductive medicine. By integrating exosome-based therapies, this research contributes to advancements in regenerative medicine and the development of effective oncofertility solutions.