Placenta-derived mesenchymal-like adherent stromal cells are isolated from full-term placentas obtained from scheduled Caesarean sections. The cells are expanded in a 3D bioreactor and have limited differentiation capacities.
The placenta is cut into pieces, washed with Hank’s buffer solution, and incubated for three hours at 37°C with 0.1% collagenase. The digested tissue is roughly filtered, washed, and seeded in culture medium in 80 cm2 flasks and incubated at 37°C in a tissue culture incubator under humidified conditions and 5% CO2. After 2-3 days, during which the cells adhere to the flask surface, cells are washed with PBS and culture medium is added. Culture medium: DMEM, D-glucose (1 g/L), fetal bovine serum (8%), gentamicin–IKA (45 μg/mL), fungizone (0.25 μg/mL), and L-glutamine (2 mM). Cells are then grown in 3D by transferring them to a bioreactor for 2-3 weeks.