Cell-free fat extract prevents diminished ovarian reserve by inhibiting granulosa cell senescence.

backgroundAge-related diminished ovarian reserve (DOR) leads to declining fertility, miscarriage, and systemic health issues. Cell-free fat extract (CEFFE) has demonstrated therapeutic effects in various tissues. In our previous study, CEFFE successfully improved ovarian function in mice without adverse effects; however whether it can prevent DOR remains unclear. This study aimed to determine if early intervention with CEFFE can delay DOR and extend reproductive lifespan, exploring its potential as a novel clinical approach for women with DOR.

methodsThe mice in the Prophylactic group received tail vein injections of 200 μL of CEFFE per mouse every 3 days for three months; the Control group were administered an equivalent volume of saline injections. Post-treatment outcomes assessed included body weight, ovarian weight, follicle count, embryo quality, production rates, and levels of serum hormones. Safety was evaluated via organ weights and hematoxylin and eosin staining in parents and offspring. The impact of CEFFE on cell proliferation, hormone synthesis, oxidative stress, and senescence was assessed via Cell counting Kit-8 assays, enzyme-linked immunosorbent assays, and reverse transcription quantitative real-time PCR, 1,1',3,3'-tetraethyl-5,5',6,6'-tetrachloroimidacarbocyanine iodide and Senescence-associated beta-galactosidase staining.

resultsCompared with the Control group, CEFFE treatment effectively preserved ovarian function in aging mice, improving ovarian weight, hormone levels, and follicular development, and reduced follicular atresia. CEFFE treatment enhanced embryo quality and development, induced a higher pregnancy rate, reduced the number of abnormal pregnancies, and increased the litter size and the number of live births. CEFFE demonstrated favorable safety in the Prophylactic group and their offspring, with no significant adverse effects on organ morphology or coefficients, except for increased liver weight in treated mice. Transcriptomic analysis suggested CEFFE influences cell proliferation, hormone response, and oxidative stress pathways in ovarian granulosa cells, which was verified in primary mouse granulosa cells. In vitro studies demonstrated that CEFFE pre-treatment alleviated the phenotype of Control mice by inhibiting oxidative stress, promoting proliferation, and enhancing hormone secretion.

conclusionsEarly prophylactic administration of CEFFE in adult mice can prevent DOR and extend their reproductive lifespan by inhibiting granulosa cell senescence.