Thawing Culture Cells

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Revision as of 14:30, 27 September 2017 by Lgunder (Talk | contribs) (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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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 and media bottle with 70%ethanol
  • Add 10mL of media to plate in sterile hood
  • Pipette cells from vile into plate with media (evenly pipette/distribute throughout plate)
  • 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