α mem medium

In the cultured groups, the fragments (N = 26 fragments in each group) were cultured individually in 96-well V-bottom culture plates for 8 days [47 (link),48 (link)]in 300 μl α-MEM medium (Biowest, Farnce) supplemented with 10% fetal bovine serum (FBS) (Gibco, UK), 50 mg/ml streptomycin, 60 IU/ml penicillin, 10 μg/ml insulin transferrin selenium (ITS) (Gibco, UK), and 0.5 IU/ml human recombinant follicle stimulating hormone (rFSH) (Merck, Germany) at 37 °C in a humidified incubator with 5% CO₂ [49 (link)]. Two-thirds of the media was replaced with fresh media every 48 h. Aspirated media of eight fragments were stored at −20 $°$ C for glucose uptake and estradiol production evaluation. After the 8-day culture period, 6 fragments were immersed in RNA lather (Yekta Tajhiz Azma, Iran) for real time PCR assessments, 10 fragments were stored at −80 °C for biochemical assays, and 10 fragments were fixed for histological assessments. In addition to the cultured groups, the procedure was applied to the fragments that were not cultured for comparison.