ASCs from human donors were commercially acquired (Stempro® Human ASCs, Thermo Fisher Scientific, Waltham, MA, USA) and characterized as previously described [21 (link)]. Myometrial cells were isolated from female donors after obtaining approval from the Institutional Review Board of the Université Catholique de Louvain (IRB reference 2020/14AOU/410). To this end, donor myometrial tissue was cut into 1 mm2 pieces using a sterile scalpel, then immersed in Dulbecco’s modified Eagle’s medium (DMEM/F12; 11320-033; Gibco), 1 µg/mL collagenase (C2674; Sigma) and 1 IU/µL DNase (D4263, Sigma). After incubation for 3 h in a warm bath at 37 °C with gentle agitation, the undigested tissue was filtered through a 100 µm cell strainer [34 (link),35 (link)].
ASCs and myometrial cells were cultured in a humidified incubator at 37 °C and 5% CO2 in 75 cm2 flasks in a basic culture medium composed of DMEM/F12 1×, 10% heat-inactivated fetal bovine serum (16140071; Gibco), and 1% antibiotic-antimycotic (15240-062; Gibco). The medium was changed every other day until 80–90% confluence was reached. All cells used were between passages 6 and 8. All the in vitro experiments were performed in triplicate.
ASCs and myometrial cells were cultured in a humidified incubator at 37 °C and 5% CO2 in 75 cm2 flasks in a basic culture medium composed of DMEM/F12 1×, 10% heat-inactivated fetal bovine serum (16140071; Gibco), and 1% antibiotic-antimycotic (15240-062; Gibco). The medium was changed every other day until 80–90% confluence was reached. All cells used were between passages 6 and 8. All the in vitro experiments were performed in triplicate.