Human preterm- and term-UC-MSCs were isolated and cultured as described previously [39 (link), 41 , 42 (link)]. Briefly, preterm- and term-UC (2–3 g wet weight) were collected, cut into 2–3 mm3 pieces, enzymatically dissociated with Liberase DH Research Grade (Roche, Mannheim, Germany), and filtered through a 100 µm cell strainer (BD Bioscience, Bedford, MA, USA). Collected cells were cultured at 37 °C (5% CO2 and 95% air) in alpha-minimum essential medium (Wako Pure Chemical, Osaka, Japan) containing 10% fetal bovine serum (Millipore Sigma, St. Louis, MO, USA) and 1% antibiotic–antimycotic solution (Invitrogen, Carlsbad, CA) until reaching confluency and then subcultured.
Human BM-MSCs purchased from Lonza (Tokyo, Japan) were cultured at 37°C (5% CO2 and 95% air) in low-glucose Dulbecco’s modified Eagle medium (Life Technologies, Carlsbad, CA, USA) containing 10% fetal bovine serum and 0.1 mg/mL kanamycin (Invitrogen) until reaching confluency and were subcultured as described previously [43 (link)].
Human BM-MSCs purchased from Lonza (Tokyo, Japan) were cultured at 37°C (5% CO2 and 95% air) in low-glucose Dulbecco’s modified Eagle medium (Life Technologies, Carlsbad, CA, USA) containing 10% fetal bovine serum and 0.1 mg/mL kanamycin (Invitrogen) until reaching confluency and were subcultured as described previously [43 (link)].
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