Difference between revisions of "Thawing Culture Cells"

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(Created page with "Materials * Media( High Glucose DMEM: Pre-prepared 500mL: 1% PSG and 10% PBS) *C2C12 cells Protocol * Thaw vile of C2C12 cells in 37C water bath approx. 1 min * Sterilize vile...")
 
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Protocol
 
Protocol
 
* Thaw vile of C2C12 cells in 37C water bath approx. 1 min
 
* Thaw vile of C2C12 cells in 37C water bath approx. 1 min
* Sterilize vile and media bottle with 70%ethanol
+
* Pipette 10mL of media to plate in sterile hood
* Add 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

Revision as of 14:36, 27 September 2017

Materials

  • Media( High Glucose DMEM: Pre-prepared 500mL: 1% PSG and 10% PBS)
  • C2C12 cells

Protocol

  • Thaw vile of C2C12 cells in 37C water bath approx. 1 min
  • Pipette 10mL of media to plate in sterile hood
  • 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
  • Incubate plate in 37C incubator
  • Check cells in 4-6 hours for growth (under microscope)
  • Replace media after significant growth approx. 85-90%

Notes: Sanitize surface, vials, hood etc with 70% ethanol and paper towel, Always return plates to incubator, Trash all cell vials and waste in contact into biohazard trash for autoclave, Review cells under microscope for growth periodically See Relevant links Splitting Cells Culturing and Differentiating C2C12 Cells