The porcine ovary is an invaluable model for human ovarian research and critical for agricultural reproductive efficiency. Traditional two-dimensional (2D) models fail to recapitulate the native ovary’s 3D structure and functions, limiting their utility. This study aimed to establish a stable 3D porcine ovarian organoid model, evaluate its utility, and explore melatonin’s regulatory effects. Porcine ovarian tissues from prepubertal gilts were digested, cultured in Matrigel® with optimized medium. Organoids were characterized via microscopy, qRT-PCR, and H while melatonin (1–20 μM) effects were tested via Cell Titer-Glo and qRT-PCR analysis. Typical round organoids formed on day 7, maintaining stability up to passage 10, expressing ovarian biomarkers ( EPACM , CDH1 , FSH , DDX4 , etc.), and mimicking in vivo histology. Lentivirus infected organoids dose-dependently. Melatonin enhanced viability/size and upregulated FOXL2 , FIGLA , and AMH genes in a dose-dependent manner. Thus, a stable and functional 3D porcine ovarian organoid model was established for gene perturbation. Melatonin benefits organoid growth via key gene upregulation, providing a novel tool for ovarian research and translational studies. • Organoids recapitulate in vivo ovarian molecular and histological features. • Lentivirus infects organoids dose-dependently, enabling gene perturbation. • Melatonin boosts organoid growth via upregulating FOXL2/FIGLA/AMH. • Model bridges agricultural and human ovarian translational research.
Hu et al. (Mon,) studied this question.
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