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In vitro  activation and growth of primordial dormant follicles to produce fertilizable oocytes would provide a useful instrument for fertility preservation. The employment of Phosphatase and TENsin homolog (PTEN) inhibitors, in combination with Protein kinase B (Akt) stimulating molecules, has been previously employed to increase follicular activation through the stimulation of the PTEN-Akt pathway.    Methods  We aim to establish improved  in vitro  activation also for cancer patients whose ovarian tissue has already been cryopreserved. Fresh and previously cryopreserved human ovarian cortex were exposed to short-term, low-concentration and ovary-specific treatment with only a PTEN inhibitor.    Results  Our  in vitro  activation protocol enhances the activation mechanisms of primordial follicles in both fresh and cryopreserved samples, and enlarges growing populations without inducing apoptosis in either follicles or the surrounding stroma. Treatment augments estradiol secretion and restores the expression levels of the previously diminished Anti-Müllerian hormone by means of cryopreservation procedures. Genomic modulation of the relative expression of  PTEN  pathway genes was found in treated samples.    Conclusion  The  in vitro  activation protocol offers new alternatives for patients with cryopreserved tissue as it increases the pool of viable activated follicles available for  in vitro  growth procedures. The combination of ovarian tissue cryopreservation and  in vitro  activation of primordial follicles, the main ovarian reserve component, will be a major advancement in fertility preservation.

plos one

Short-Term PTEN Inhibition Improves In Vitro Activation of Primordial Follicles, Preserves Follicular Viability, and Restores AMH Levels in Cryopreserved Ovarian Tissue From Cancer Patients