Patient-derived decidual organoids reveal rhG-CSF-associated protective responses involving JAK2/STAT3 signaling in recurrent spontaneous abortion.

BACKGROUND

Recurrent spontaneous abortion (RSA) is a complex reproductive disorder frequently lacks effective therapeutic interventions. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) has shown promise in improving pregnancy outcomes in RSA, but its underlying mechanisms remain unclear. Organoid technology offers new opportunities for disease modeling and mechanistic research in reproductive medicine.

METHODS

Decidual tissue samples from RSA patients were utilized to establish decidual organoids using three-dimensional (3D) culture techniques. These organoids were characterized via morphological observation, hematoxylin and eosin (H&E) and Periodic Acid-Schiff (PAS) staining, immunofluorescence, and immunohistochemistry to validate key marker expression and hormone responsiveness. The effects of rhG-CSF on organoid viability were evaluated using luminescence assays. Transcriptomic sequencing and bioinformatics analyses were performed to identify differentially expressed genes and pathways following rhG-CSF stimulation. Protein expression and signaling pathway activation were further assessed via Western blot analysis.

RESULTS

Decidual organoids were successfully established and recapitulated the structural and functional features of primary tissue, including hormone responsiveness and marker expression. rhG-CSF significantly promoted organoid proliferation, enhanced anti-apoptotic and angiogenic capacity, and induced inflammatory responses. Transcriptomic and pathway analyses indicated an enrichment and activation of the JAK2/STAT3 signaling pathway following rhG-CSF treatment. Western blot indicated increased phosphorylation of JAK2 and STAT3, upregulation of PCNA, VEGFA, IL-1β, IL-6, and Bcl-2, and downregulation of Bax in response to rhG-CSF.

CONCLUSION

A patient-derived decidual organoid model was established and validated to recapitulate in vivo tissue characteristics. In this in vitro RSA model, rhG-CSF appeared to exert protective effects by promoting proliferation, inhibiting apoptosis, and enhancing angiogenesis-responses that are potentially associated with the activation of JAK2/STAT3 signaling. These findings provide new insights and experimental evidence that may inform future RSA treatment strategies.