Umbilical cord blood-derived mesenchymal stem cells (Cedars-Sinai Medical Center)

Umbilical cord blood (UCB) samples are collected within 10 minutes of delivery, after removal of the placenta. UCB samples are subjected to Histopaque separation and the interface layer is then collected and centrifuged at 500g for 10 minutes. Cells are washed and further centrifuged at 350g for 5 minutes. UCB mononuclear cells (MNCs) are seeded in fibronectin-coated culture dishes, and cultured in Dulbecco’s modified Eagle medium (DMEM) high-glucose medium supplemented with fetal bovine serum (FBS, 10%), and penicillin-streptomycin (1%). Cells are incubated for 5-7 days in a humidified atmosphere containing 5% CO2, before cultured medium is replaced and non-adherent cells are removed.