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BACKGROUND
Culturing cells and its maintenance is as important as cell growth control. Cells grow fast and there will be so many plates after many passages. We have to preserve cells for further use in future, for that, it is important to preserve healthy and integrated cells. It saves all expensive chemical resources for the cell culture also.
The aim of this experiment is to preserve healthy and integrated cells for future application.
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REQUIREMENTS
Sample: Animal cells in a culture plate (confluent stage).
Apparatus: Centrifuge tube – 15 ml
Serological pipette
Centrifuge with rotor
Chemicals: 1X HBSS
Cell culture media
0.5% trypsin solution
Serum
DMSO
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PROCEDURE
Freezing media:
|
Content |
Volume |
Composition |
|
Serum DMSO Cell culture media |
500µl 100µl 400µl |
50% 10% 40% |
Get the confluent cell plate and wash with 1X HBSS and add 5 ml 0.5% trypsin solution (per 60 mm petri dish), incubate at 37°C for 3-5 min and make sure that all the cells are detached from the plate. Add 5-7 ml of cell culture media and gently mix it and try to dissociate all the cell clumps in the solution. Centrifuge it at 300x g for 3-5 min and discard the supernatant further add the freezing media and mix properly and immediate keep in to the -20°C for 1-2 hrs then you can transfer in to liquid nitrogen tank for long time storage.1
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CONCLUSION
In this experiment, we performed freezing live cells for the future application in liquid nitrogen tank.
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REFERENCES
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Kumar A, Mishra HK, Dwivedi P, Subramaniam JR. Secreted trophic factors of Human umbilical cord stromal cells induce differentiation and neurite extension through PI3K and independent of cAMP pathway. Ann Neurosci. 2015;22(2):97-106.
