Thawing frozen A1 cells: Difference between revisions
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== Materials | == Materials == | ||
* Gelatin (warmed to 37ºC) | * Gelatin (warmed to 37ºC) | ||
* HS for A1 cells | * HS for A1 cells | ||
| Line 7: | Line 7: | ||
* frozen cells into liquid nitrogen | * frozen cells into liquid nitrogen | ||
== Procedure | == Procedure == | ||
* put 10 ml Gelatin into the new 175 cm2 flasks (remove the remaining liquid) | * put 10 ml Gelatin into the new 175 cm2 flasks (remove the remaining liquid) | ||
* pipette 5 ml fresh media into the 15 ml FALCON tube | * pipette 5 ml fresh media into the 15 ml FALCON tube | ||
Revision as of 09:23, 25 March 2015
Materials
- Gelatin (warmed to 37ºC)
- HS for A1 cells
- 1x 15 ml FALCON tubes
- Pipettes
- 1x 175 cm2 flasks
- frozen cells into liquid nitrogen
Procedure
- put 10 ml Gelatin into the new 175 cm2 flasks (remove the remaining liquid)
- pipette 5 ml fresh media into the 15 ml FALCON tube
- thaw the cells into a baker contains warm (nearly 25ºC) water until there is a very small piece of ice left
- transfer the cells into the FALCON tube
- centrifuge at 1000 rpm, 3 min at 4ºC
- aspirate the supernatant
- dissolve the pellet into 1 ml fresh media
- fill in the new flasks 25 ml HS for A1 cells
- transfer the cells in the flask
- label the flask with the name of the cells, the passage number and the date
- check the cells the next day, that they have attached on the flask
The cells grow at 25ºC, 2% CO2