Hi,
I applied on PC12 cells 100 ng/ml NGF and incubated for 72 hours to differentiation. After 72 h, approximately most cells differentiate and then I applied an anticancer drug but all of the cells including the control group died. The medium I used while applying the drug is the same as the medium I used for differentiation. I couldn't find my mistake. I'm waiting your idea or advice.
By control group, do you mean cells grown the same condition but without the cancer drug?
You need a no drug control under the same conditions as the drug but without drug. And yes the control cell must have high survival.
It is possible that extended serum starvation induces apoptosis in these cells. It is critical that you know the survival tendencies including plating density and resistance to serum starvation induced apoptosis.
Understand that serum "starvation" often refers to the time since fresh serum (media change).
Cells are supposed to die without serum. The more "normal" a cell line is, the more sensitive it is to serum starvation.
72hours with no fresh serum in many cell lines increases apoptosis without any other treatment.
Hello,
What is the differentiating medium you are using. It should consists of basal medium (DMEM) + 1% horse serum +1% penstrep + 100ng/ml NGF.
Have you added minimum quantity of serum to the differentiating medium? It is very much needed for cell survival. This could be one of the reason for cell death.
Best,
Dear Malcolm Nobre
I used RPMI+%1HS+%1PS and 100 ng/ml NGF to differentation. After 72 hours incubation, I saw differentiation in cells very clearly. My problem is to keep the cells alive after the cells differentiate. Should i use RPMI+%1HS+%1FBS+%1PS to keep alive after differentiation? I dont know.
Neal Edward Beeman ,
Yes control group is cells grown the same condition but without the cancer drug.
When you say serum do you mean FBS?
Yes I do refer to FBS as serum.
Be careful of all of the language here. Some researchers refer to "serum starvation" as the last time media was refreshed. Other researchers pipette away the media and replace with a serum free media.
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