Shreffler:CFSE Labeling

Overview

CFSE is used to fluorescently label live cells in order to track them. 5,6-CFDA/SE is membrane permeable, but trapped in the cell when converted by intracellular esterases. CFSE is equally partitioned to daughter cells during division and can be used to measure cell proliferation.

Materials

• 15 mL polypropylene tubes
• PBS
• 5,6-CFDA/SE Invitrogen C1157 dissolved in DMSO at 5 mM and stored at -20°C.

Procedure

1. Suspend PBMCs at 10x10⁶ cells/mL in PBS alone.
2. Cells should be in PBS for CFSE labeling.
3. If CFSE negative control is needed, remove cells now.
4. Make '2X' concentration (10 μM) in PBS
   • For example: add 5 mL PBS + 10 μL 5 mM CFSE
5. Combine 1:1 PBMC cells + CFSE (e.g. 5 mL PBMCs + 5 mL CFSE) in 15mL tube.
6. Place in 37°C H₂O bath x 10 min.
7. Wash in 10 mL complete medium.
8. Resuspend in medium at desired density

Notes

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References

1. =8176234