Hi everyone,
Recently I have been performing ki67 proliferation assay to determine the effects of TGFbeta1 on fibroblasts. My supervisor told me that I need to starve the fibroblasts for at least 12 hours before I add the TGF beta1 treatment. However, I am confused why I need to starve the cells as this is an experiment detecting cell proliferation. I've been searching for papers in this field. However, no paper illustrates whether the authors starved the cells or not. Hence, can anyone could help me clarify this question?
Thank you very much in advance!
Hello Aries Huang
Your supervisor is right. Serum starvation is widely used for synchronizing cells by arresting them in the G0/G1 phase of the cell cycle. In this way, cells at different stages of the cell cycle in a culture are brought to the same phase.
If you are still confused, you may have three experimental set-ups.
1. You may starve the cells 24 hours after plating without serum for 12 hours before TGF beta1 treatment, after which the starvation medium may be changed to normal medium containing TGF beta1.
2. You may treat the cells with TGF beta1 24 hours after plating with normal medium containing TGF beta1.
3. In the third experimental set-up, you may starve the cells 24 hours after plating without serum for 12 hours before TGF beta1 treatment, after which you may treat the cells with TGF beta1 but maintain the starvation condition during the treatment.
Best.
Hi Malcolm Nobre , thank you so much for your information. Can I ask one more question, if I set up the experiments as you mentioned, what results should I expected? Thanks a lot in advance!
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