(310) Rejuvenation and Functional Recovery of the Ovary through Human Umbilical Cord Mesenchymal Stem Cells in a Mouse Model of Cyclophosphamide-Induced Premature Ovarian Insufficiency

Jeong Eun Lee¹, Ji Won Lim², Han Seul Yang², Ji Hyang Kim^1*

¹ Fertility Center of CHA Bundang Medical Center, College of Medicine, CHA University, Seongnam, Republic of Korea

² Department of Biological Sciences, KAIST Stem Cell Center, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea

Abstract Text:

Introduction: Premature Ovarian Insufficiency (POI) represents a significant reproductive challenge with limited therapeutic options. This study investigates the regenerative potential of human umbilical cord mesenchymal stem cells (hUC-MSCs) in a cyclophosphamide (CP)-induced POI mouse model.

Methods: Following CP administration (200 mg/kg), female ICR mice received varying doses of hUC-MSCs (low: 5.33×10⁴, moderate: 8×10⁴, or high: 1.07×10⁵ cells/mouse) via tail vein injection. Ovarian function was assessed through histological analysis, apoptosis assays, AMH immunostaining, in vitro fertilization outcomes, and single-cell RNA sequencing.

Results: Histologically, hUC-MSCs restored ovarian architecture and follicular populations in a dose-dependent manner, with a 77% increase in primordial follicles in the high-dose group. TUNEL assays demonstrated significant reduction in apoptotic cells, while AMH immunostaining showed restored expression patterns in granulosa cells. In vitro fertilization outcomes exhibited substantial enhancement, with high-dose treatment restoring fertilization (73.4%) and blastocyst development rates (73.9%) to near-control levels. Single-cell RNA sequencing identified upregulation of wound healing and extracellular matrix remodeling pathways in epithelial cells (Col3a1, Col1a2), enhanced intercellular communication networks, and VEGF-related angiogenic pathways in stromal cells.

Conclusion: hUC-MSCs therapy effectively reverses CP-induced ovarian damage through multiple regenerative mechanisms.

Discussion: This study provides unprecedented insight into hUC-MSCs' multi-faceted therapeutic mechanisms in ovarian regeneration, offering promising implications for treating chemotherapy-induced POI and advancing fertility preservation strategies.