CELL CULTURE PROTOCOLS
CELL SPLITTING (FOR 10cm Flask) (Amount of media – 10ml)
Before the start of cell culture work -
A. Remember to warm the cell culture media in 37ᴼC for 30 min.
B. Clean the Hood with 70% ethanol
C. Cell Culture works should always be done by wearing gloves.
1. Allow the cells to reach a confluency of 90%.
After the cells reach the desired confluency
2. Aspirate the media of the cell
3. Wash the cells with 5ml PBS
4. Add 1ml of trypsin to the cell and incubate the plate in 37ᴼC for 5 minutes.
5. Check under the microscope whether the cells have detached. (You can additionally tap the sides of the flask with your hand and ensure that the lumps of cells are dispersed).
6. After the cells detach, add 9ml new media immediately to dilute the trypsin
7. Resuspend the detached cells in the new media by pipetting
8. Now add the desired amount cells (From step 6) with the new media into new plates – According to the concentration of cells you want.
CELL SPLITTING (FOR 10cm Flask) (Amount of media – 10ml)
Before the start of cell culture work -
A. Remember to warm the cell culture media in 37ᴼC for 30 min.
B. Clean the Hood with 70% ethanol
C. Cell Culture works should always be done by wearing gloves.
1. Allow the cells to reach a confluency of 90%.
After the cells reach the desired confluency
2. Aspirate the media of the cell
3. Wash the cells with 5ml PBS
4. Add 1ml of trypsin to the cell and incubate the plate in 37ᴼC for 5 minutes.
5. Check under the microscope whether the cells have detached. (You can additionally tap the sides of the flask with your hand and ensure that the lumps of cells are dispersed).
6. After the cells detach, add 9ml new media immediately to dilute the trypsin
7. Resuspend the detached cells in the new media by pipetting
8. Now add the desired amount cells (From step 6) with the new media into new plates – According to the concentration of cells you want.
