Mesenchymal Stem Cells Culturing

  1. Mesenchymal stem cells from human umbilical cord were provided by Prof. Chun-Min Lo from National Yang Ming University.
  2. View cultures using a microscopy and after the cultures was full of the dish, remove the spent medium.
  3. Wash the cell monolayer with PBS and remove it.
  4. Add HyQTase(1ml per 25 cm2 of surface area) and incubate for 1 minutes.
  5. After making sure cells were detached, resuspend the cells with medium(more than 5 folds volume of HyQTase).
  6. Centrifuge under 2000 rpm, 5minutes, 25°C.
  7. Remove the supernatant and add 5 ml medium to suspend cells.
  8. Count the cells by Bright Line Counting Chamber. Subject the cell diversity to 104/ml
  9. Seed the cells for 104/well, 1ml/well in the 24-well plate.
  10. Until cells grow full of each well, do the coculturing experiment.