Dmem f12 primary growth medium

Primary genital epithelial cells were isolated from cervical and endometrial tissues obtained from women aged 30–59 years (mean age 42.9 ± 7.2) undergoing hysterectomies for benign gynecological reasons at Hamilton Health Sciences Hospital. Informed written consent was obtained in accordance with the approval of the Hamilton Integrated Human Research Ethics Board. A detailed protocol for the isolation and culture of primary GECs has previously been described by Kaushic et al., [26 (link)]. Briefly, endometrial or endocervical tissues were minced into small pieces, digested in an enzyme mixture for an hour at 37 °C. GECs were isolated by a series of separations through nylon mesh filters (Small Parts, Miramar, FL, USA), resuspended in DMEM/F12 primary growth medium (Invitrogen, Burlington, ON, Canada), and seeded onto Matrigel-coated (BD Biosciences, Mississauga, ON, Canada) tissue culture inserts (BD Biosciences). GEC cultures were grown for 5–7 days until confluent monolayers were formed. The confluency was monitored by transepithelial resistance (TER) measured by a volt ohm meter (EVOM; World Precision Instruments, Sarasota, FL, USA). Confluent monolayers showing TER values greater than 1000 Ω/cm² were used for further experiments. The purity of epithelial monolayers was between 95% and 98%, with no trace of any hematopoietic cells.