The ovary tissue of patients (n = 11) of non-ovarian factors containing primordial follicles were collected using excision surgery at the Department of Obstetrics and Gynecology in Shanghai Changzheng Hospital. All the participants are transsexual patients that had normal ovarian function and were not under hormonal treatment before the surgery. This study is approved by the Institutional Medical and Ethical Review Committee of Second Military Medical University. Clinical characteristics of cases were shown in Table 1. After being cut into 1 mm3 cubes, cortical tissues were placed on culture plate inserts (Millipore) and cultured in 400 μl of DMEM/F12 (Gibco) containing 10% human serum albumin (CSL Behring), 1% penicillin-streptomycin solution (HyClone), 0.3 IU/ml FSH, and 0.05 mg/ml L-ascorbic acid under a membrane insert to cover ovaries with a thin layer of medium. Ovaries were treated with 1–20 μM MHY1485 (10 mM in 1 ml DMSO, APExBIO, United States) and cultured for 3 h before immunoblotting analysis. Control ovaries were treated with solvent only. Other pairs of ovaries were cultured for 2 days with a medium change after 24 h of culture. Ovaries were fixed with formalin for histological analysis or prepared for transplantation before weighing.

Clinical characteristics of transsexual patients.

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