Cell freezing is done if the cell type used is rare or for storage purposes or for transport. It is a very simple procedure, only require a few steps.
Protocol (For freezing 1ml)
1. Trypsinize cells
2. Count cells
3. Resuspend cells with medium (Cell density should be ~50 000 - 1 000 000 cells/ml)
4. Add 900ul of cell suspension and 100ul of (Dimethyl Sulfoxide)DMSO into a cryovial.
5. Freeze at -80 degree celsius or -196 degree celsius.
Note: DMSO acts like a cryoprotectant which prevents damage arising from freezing.
Alvin
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Hi Alvin,
ReplyDeleteMay I know why the cells has to be frozen at two specific temperatures, -80 and -196 degress celsius, and not any other temperature in between the above mentioned temperatures?
Regards,
Zhu Zhijie Dennis
0700847G
TG01
Hello Alvin,
ReplyDeleteI would like to know why do we require a cell density of 50 000 - 1 000 000 cells/ml? Is it to make sure that there would still be the presence of cells after transportation?
If so, may i also ask what is the viability of these cells that are transported or stored? Would most of them survive?
THANKS!
Renee
TG02
0703634F
Hi Alvin ^^
ReplyDeleteJust wanna check with u, beside DMSO, is there any other cryoprotectant which can be used for cell freezing?
Vo Thu Hong Anh [Jess]
0705364H
Alvin,
ReplyDelete(1) What is the difference between your freezing medium and the commercial freezing medium e.g. Invitrogen Recovery Freezing Medium?
(2) What does your medium contain?
LI Yinliang Alex 0704894E
TG02 Group 2
29 September 2009
hihi,
ReplyDeletei would like to know if you count the cells when they are resuspend in trypsin or you resuspend in other reagent.
thank you.=)
Yeo Sok Kian Jocelyn
0703359J
Thanks for your information and putting the important article.I would like to tell you that you have given me much knowledge about it.
ReplyDeleteKetone Body Assay Kit