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Latest revision as of 18:11, 25 September 2012
Protocol
Mesenchymal Stem Cells Culturing
- Mesenchymal stem cells from human umbilical cord were provided by Prof. Chun-Min Lo from National Yang Ming University.
- View cultures using a microscopy and after the cultures was full of the dish, remove the spent medium.
- Wash the cell monolayer with PBS and remove it.
- Add HyQTase(1ml per 25 cm2 of surface area) and incubate for 1 minutes.
- After making sure cells were detached, resuspend the cells with medium(more than 5 folds volume of HyQTase).
- Centrifuge under 2000 rpm, 5minutes, 25°C.
- Remove the supernatant and add 5 ml medium to suspend cells.
- Count the cells by Bright Line Counting Chamber. Subject the cell diversity to 104/ml
- Seed the cells for 104/well, 1ml/well in the 24-well plate.
- Until cells grow full of each well, do the coculturing experiment.
