Difference between revisions of "Thawing Culture Cells"
From Bridges Lab Protocols
(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, | ||
Latest revision as of 15:05, 4 October 2017
Materials
- 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 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)
- 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
- Passage cells normally, see Splitting Cells.
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
