Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) were isolated from the Wharton's jelly (WJ) tissue of human umbilical cords. WJ-derived cells have a fibroblast-like morphology and a population doubling time (PDT) of 23 hours. WJ-MSCs express mesenchymal antigens and lack expression of hematopoietic markers. Cells can undergo adipogenic, osteogenic and chondrogenic differentiation.
Isolated Wharton's jelly (WJ) was cut into small pieces and plated in culture medium comprised of Dulbecco’s modified Eagle’s medium low-glucose (DMEM-LG) supplemented with fetal bovine serum (FBS, 10%), L-glutamine (2 mM), penicillin (100 U/ml), streptomycin (100 μg/ml), Fungizone (25 μg/ml), basic fibroblast growth factor (bFGF, 5 ng/ml) and epidermal growth factor (EGF, 5 ng/ml). The first medium change was after 4 days, followed by medium changes every 3-4 days until cells were confluent. Cells were passaged, using trypsin with diamine tetraacetic acid (EDTA), and maintained in culture medium.