Differentiation of Human Umbilical Cord Mesenchymal Stem Cells Into Pre‐Granulosa Cells and Further Maturation

Mesenchymal stem cells (MSCs) are adult stem cells with the capacity to differentiate into several cell types, including hepatocyte-like cells, neural-like cells, islet-like clusters and so on. However, the differentiation into ovarian-related cells such as ovarian granulosa cells (OGCs) has not been well studied. Here, we established an efficient culture system to differentiate human umbilical cord mesenchymal stem cells (HUCMSCs) into pre-granulosa cells, which can further mature into granulosa-like cells. HUCMSCs were cocultured with hormones (E2, FSH) and cytokines (TGF-β), a simple differentiation method was used to induce morphological changes and positive expression of forkhead transcription factor (FOXL2). In the process of further mature, the cells differentiated into mature granulosa-like cells, expressed mature granulosa cell markers (FSHR, AMHR2, CYP19A1) and likely possesses some granulosa cell hormone secretion capacity. In conclusion, we successfully established an efficient protocol to generate pre-granulosa cells from HUCMSCs that can further differentiate into mature granulosa-like cells, opening a new avenue for the further study of therapies for female reproductive health.