Thawing frozen A1 cells: Difference between revisions
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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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== | == Material == | ||
* 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'' | ||
Latest revision as of 14:20, 8 June 2015
Material
- 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