Difference between revisions of "Thawing frozen A1 cells"
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(Created page with "== 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: == - p...") |
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| Line 1: | Line 1: | ||
== Materials: == | == 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: == | == 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 | The cells grow at 25ºC, 2% CO2 | ||
Revision as of 07:43, 24 September 2014
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