Wharton-Jelly-derived mesenchymal stem cells (Qingdao University)

The umbilical cord is washed twice with phosphate buffered saline (PBS) and then dissected with scissors into pieces (≈1cm³). These are plated in a cell culture dish containing low glucose Dulbecco`s Modified Eagle`s Medium medium supplemented with non-animal-derived serum (5%). Cells are maintained in a humidified atmosphere with 5% CO₂ at 37°C. After 3 days of culture, the medium is replaced to remove the tissue and non-adherent cells, and changed twice weekly thereafter. Once 80% confluence is reached, the adherent cells (passage 0) are detached with trypsin (0.125%) and passaged in the cell culture dish.