MenSC culture

Three menstrual blood donors were diagnosed without reproductive system diseases, aged 25, 25, and 27 years old. All donors gave consent, and all procedures were approved by the Ethics Committee of Shengjing Hospital affiliated to China Medical University (2017PS330K).

The culture and identification of MenSCs were performed in the same way as our previous article [17]. Briefly, menstrual blood was sterilely collected and isolated. The cells were regularly cultured in DMEM/F12 medium (1:1, HyClone, Logan, UT, USA) with 10% fetal bovine serum (Gibco, Waltham, MA, USA ) under 37 °C in an atmosphere containing 5% CO₂. The culture medium was changed every 3 days until adherent cells reach 80–90% confluence, then the cells were passaged by 0.25% trypsin (Sigma-Aldrich). P3 MenSCs were collected and measured surface markers of mesenchymal stem cell. MenSCs were transplanted before the sixth passage.