Female fertility Female fertility
Conditions
Interventions
Not applicable.
Sponsors
Erasmus MC, Universitair Medisch Centrum Rotterdam
Eligibility
Inclusion criteria
Inclusion criteria: Underwent fertility preservation procedures.Ovarian tissue collected prior to any exposure to chemotherapy.Provided consent for the use of surplus tissue for research.
Exclusion criteria
Exclusion criteria: Tissue shows significant degradation.Tissue was collected after exposure to chemotherapy or other medical treatments.The individual displayed comorbidities or non-cancer-related conditions that may affect granulosa cell function or ovarian development.
Design outcomes
Primary
| Measure | Time frame |
|---|---|
| Transcriptomic profile follicles and surrounding tissue. We will characterize the transcriptomic landscape of cryopreserved ovarian tissue using scRNA sequencing. This analysis will include all cells present in the tissue pieces. The resulting cell-specific transcriptional profiles will reflect the physiological state of each cell type and provide a comprehensive overview of gene expression across the tissue. Epigenetic profile follicles and surrounding tissue. We will analyze the epigenetic landscape of cryopreserved ovarian tissue using single-cell MeD-seq, a technology that enables genome-wide DNA methylation profiling at single-cell resolution. By integrating methylation data with corresponding single-cell transcriptomic profiles, we aim to uncover regulatory relationships within specific cell types. This combined dataset will serve as a reference framework for defining the epigenetic and transcriptional identity of GrCs, guiding the refinement of our in vitro differentiation protocols.iPSC induction Successful reprogramming of purified GrCs into iPSCs using a non-integrative Sendai virus-based method will be assessed. Reprogramming efficiency will be evaluated by the emergence of colonies with pluripotent morphology and confirmed by expression of key pluripotency markers. | — |
Secondary
| Measure | Time frame |
|---|---|
| Secondary outcome parameters include the comparative analysis of the single-cell transcriptomic profiles with publicly available sequencing datasets of primary ovarian tissue to validate and contextualize the identified cell populations and gene expression signatures. Where possible, similar comparisons will be made for the epigenetic (DNA methylation) profiles, although reference datasets remain limited. Additionally, induced iPSC lines will be evaluated for their differentiation potential using standard tri-lineage differentiation assays. Their capacity to generate GrC-like cells will be tested using directed differentiation protocols, and outcomes will be benchmarked against transcriptomic and epigenetic profiles generated in this study alongside publicly available datasets. | — |
Countries
Netherlands
Contacts
Public ContactE.B. Baart
Erasmus MC, Universitair Medisch Centrum Rotterdam
Outcome results
None listed