Inactivated/activated FBS - is it critical for healthy cells (Dec/08/2005 )
For HeLa cells, what is recommended ....using inactivated 10% FBS or regulr FBS
What happens if inactivated FBS is used during culture ??
Thanks for the help.
jhil
You should for any cell line use inactivated FBS.
(inactivation is done to inactivate the complement system of the immune system that is present in the serum and which would otherwise kill your cells).
hi
i don't agree.
Inactiveted serum( if it's same as decomplemented as by 56° 30') destroys complement's protein C1 to C9 proteins but alos degradesome essential factors present in the serum.
By talking with a colleague who deals on immunology, decomplemented serum should be used in immunlogical studies. In the other studies full serum seems to be required to get conditions as close as possible as the ex vivo conditions.
i've tested transfection under decomplemented serum and results are worse.
in the other hand, some infection procedures should be done in decomplemented serum.
fred
I am growing cells (HeLa) in medium containing inactivated (56 deg;30') and so far they r growing well.
But I still have to do transfection, and I am confused what to use finally?
if any one has used these cells for transfection, please let me know your views.
thanks,
jhil
i don't agree.
Inactiveted serum( if it's same as decomplemented as by 56° 30') destroys complement's protein C1 to C9 proteins but alos degradesome essential factors present in the serum.
...
i've tested transfection under decomplemented serum and results are worse.
If you remember the other topic concerning transfection, I have always done mine in inactivated, but if it's really better to do so in non-inactivated, I'll try that.
as far as i am aware it is generally a wasteof time to inactivate the FBS we did this in my lab for yrs and then decided it didnt make any difference and stopped we just found that it was necessary to spin the aliquots in falcon tubes to remove particulate matter.
as for what to use for transfection it generally depends on what transfection reagent ur using ie fugene needs serum staring and transfectene doesnt.
hope this helps
Regards
sorry i would not be so drastic. I've done experiments in plasmid transfection in 293 and HeLa cells and i got better results in full serum. I didn't forget your posts. But as full serum is, as i know and please correct me if i'm wrong, used in routine by ajority of labs, i think that setting up a transfection assay should be done in closer conditions as routinely used.
fr sure, if results with certain cell lines and or certains transfection agents are better with inactivated serum ok use it.
I'm just gonna split these cells for 2 weeks in non-inactivated serum, and then do transfection. I'll keep a batch that I split with inactivated serum, and then compare the two...