Dmem f12

GMMe cells obtained from the American Type Culture Collection (ATCC, Manassas, VA, CRL-267) were grown in Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; Caisson Labs, Smithfield, UT, DFL14-500ML) with 10% FBS (Fetal Bovine Serum; Caisson Labs, Smithfield, UT, FBL02-500ML) at 37°C and 5% CO₂. After 48 h of growth, the cells were trypsinized, counted and prepared for RNA extraction.

Human adult-type granulosa cell tumour-derived KGN cells (Riken, Tsukuba, Japan), 293T human embryonic kidney cells (ATCC, Manassas, VA, USA), human juvenile granulosa cell tumour-derived COV434 cells (Sigma-Aldrich), HeLa human cervical cancer cells (Korean Cell Line Bank, Seoul, Korea), and A549 human lung adenocarcinoma cells (ATCC) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Caisson, North Logan, UT, USA) and DMEM/F12 (Caisson) containing 10% foetal bovine serum (FBS) and 1% penicillin-streptomycin (Caisson) at 37 °C in 5% CO₂. MRC-5 human lung normal cells (Korean Cell Line Bank) were cultured in Minimum Essential Medium containing 25 mM HEPES, 25 mM NaHCO3, 10% FBS, and 1% penicillin-streptomycin. The human B-lymphocyte cells line (GM01029; Coriell Institute, Camden, NJ, USA) were cultured in RPMI 1640 containing 15% FBS and 1% penicillin-streptomycin. Cells were transfected with Lipofectamine 2000 or 3000 (Invitrogen) according to the manufacturer’s instructions.

The Eker rat-derived uterine leiomyoma (ELT3) cell line and primary human uterine smooth muscle (UtSMC) cells were kindly provided by Dr. Lin-Hung Wei (Department of Oncology, National Taiwan University Hospital, Taipei, Taiwan). Both ELT3 and UtSMC were maintained in Dulbecco’s Modified Eagle Medium/Ham’s F-12 Medium 1:1 (DMEM/F12, CAISSON Labs, Smithfield, UT, USA) supplemented with 10% fetal bovine serum (FBS; Biological Industries, Cromwell, CT, USA), 100 units/mL penicillin, 100 μg/mL streptomycin, sodium bicarbonate (2.438 g/L), and HEPES (5.986 g/L) in a humidified incubator (37 °C, 5% CO₂).

The Eker rat-derived uterine leiomyoma ELT3 cell line was provided by Dr. Lin-Hung Wei (Department of Oncology, National Taiwan University Hospital, Taipei, Taiwan). The primary human uterine smooth muscle (UtSMC) cells were purchased from PromoCell (Heidelberg, Germany). UtSMC cells were isolated from the middle layer of the uterine myometrium and the short-tandem repeat DNA profiling was analyzed. Both cells were grown in Dulbecco’s Modified Eagle Medium/Ham’s F-12 Medium in a 1:1 ratio (DMEM/F-12; CAISSON Labs, Smithfield, UT, USA), supplemented with 10% fetal bovine serum (FBS; GIBCO, Grand Island, NY, USA), 100 units/mL penicillin (CORNING; Manassas, VA, USA), 100 μg/mL streptomycin, sodium bicarbonate (2.438 g/L, BioShop, Burlington, ON, Canada), and 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES; 5.986 g/L; BioShop) in a humidified incubator (37 °C, 5% CO₂). After resuscitation, ELT3 cells were sub-cultured with less than 20 passages, and UtSMC from 5–12 were used.