Isolation of Mesenchymal Stem Cells from Fallopian Tubes

MSCs were isolated from healthy fallopian tubes (hFTs) collected in DMEM/Hams F-12 medium (Invitrogen, Carlsbad, CA) and kept at 4 °C for processing within 24 h as described previously [15] (link). Briefly, hFTs were opened, washed twice in PBS (Gibco, Invitrogen, Carlsbad, CA) and incubated at 37 °C for 30 min in a 50 mL Falcon tube containing 5 mL of pure TrypLE™ Express (Invitrogen, Carlsbad, CA) with shaking. The supernatant was removed, washed once with 7 mL of advanced DMEM/Hams F12 in a 15 mL Falcon tube and centrifuged at 1000 rpm for 5 min at room temperature. The cells were then plated in advanced DMEM/Hams F12 (12 mL) supplemented with penicillin/streptomycin (100 µg/mL) in 75 cm³ polystyrene culture flasks and incubated at 37 °C in a humidified atmosphere containing 5.0% CO₂ (Figure 1).