Mesenchymal stem cells were isolated from umbilical cord blood samples. Cells grow in focal patches, display a spindle-shaped large cell body, express specific mesenchymal stem cell markers, and have adipogenic, chondrogenic, and osteogenic differentiation capacities. The type 2 cell type has a high chondrogenic differentiation capacity, and a low adipogenic differentiation capacity, compared to type 1 umbilical cord blood-derived mesenchymal stem cells. Type 2 cells have a longer doubling time compared to type 1 cells. Cells have a normal 46, XY karyotype.
Umbilical cord blood samples were layered on Histopaque and centrifuged for gradient separation. The interface layer was extracted, diluted and further centrifuged to collect the cells. Cells were washed, centrifuged again, counted and seeded in fibronectin-coated plates or in flasks. Cells were cultured in high-glucose Dulbecco’s modified Eagle medium (DMEM) supplemented with fetal bovine serum (FBS, 10%) and penicillin-streptomycin (1%).