Thawing Culture Cells: Difference between revisions
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 | ==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. | ||
* | * Frozen cell aliquot in Liquid Nitrogen | ||
Protocol | |||
* Thaw | ==Protocol== | ||
* | * Thaw vial of 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) | * 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 | * 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, | ||