Umbilical cord blood-derived multipotent stem cells (University of Illinois at Chicago)

Human umbilical cord blood samples (50–100 ml/unit) were obtained from healthy donors. Mononuclear cells were isolated using Ficoll–Hypaque, followed by removal of red blood cells using cell lysis buffer. Mononuclear cells were seeded into 8-well chamber slides at a density of 1–2×10⁵ cells/ml, in a total volume of 0.5 ml/well.  Alternatively, cells were seeded into 150×15 mm Petri dishes at a density of 1×10⁶ cells/ml, in a total volume of 25 ml/dish in RPMI medium supplemented with fetal bovine serum (7%) and incubated at 37ºC and 8% CO₂.