Thawing Culture Cells: Difference between revisions

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Materials
==Materials==
* Media( High Glucose DMEM: Pre-prepared 500mL: 1% PSG and 10% PBS)
* Media (Typically High Glucose DMEM: Pre-prepared 500mL: 1% PSG and 10% PBS).  May be different for different cell lines.
*C2C12 cells
* Frozen cell aliquot in Liquid Nitrogen
Protocol
 
* Thaw vile of C2C12 cells in 37C water bath approx. 1 min
==Protocol==
* Sterilize vile and media bottle with 70%ethanol
* Thaw vial of cells in 37C water bath approx. 1 min
* Add 10mL of media to plate in sterile hood
* Pipette 10mL of media to plate in sterile hood
* Pipette cells from vile into plate with media (evenly pipette/distribute throughout plate)
* Pipette cells from vile into plate with media (evenly pipette/distribute throughout plate)
* Lightly swirl the plate until bottom of plate is covered
* Review cells under microscope
* Review cells under microscope
* Incubate plate in 37C incubator
* Incubate plate in 37C incubator
* Check cells in 4-6 hours for growth (under microscope)
* Check cells in 4-6 hours for growth (under microscope)
* Replace media after significant growth approx. 85-90%
* Replace media  
* Passage cells normally, see [[Splitting Cells]].
 
Notes:
Notes:
Sanitize surface, vials, hood etc with 70% ethanol and paper towel,
Sanitize surface, vials, hood etc with 70% ethanol and paper towel,