I'm working on a wound healing project and currently trying to see whether my extract has positive effect on cell proliferation. The two cell lines that I'm using are HaCaT keratinocyte and 3T3 fibroblast. After performing the experiment for several times, I found significant positive effect on HaCaT but not 3T3. Is there any explanation for this?
My method is as follow:
1) Seed about 5,000 cells per well for HaCaT and 3,000 cells per well for 3T3
2) Incubate for 24 hours
3) Serum starve with media containing 0.1% FBS for 2 hours
4) Remove media and add new media containing 1% FBS with extracts
5) Incubate for another 24 hours
6) Perform MTT cell viability assay
By the way, I'm using epithelial growth factor (EGF) as positive control and both cell lines show positive cell proliferation effect.
Extract only works on some cells, but not all. Just like some cytotoxicity assay, some of the plant extract can suppress the growth of certain cell lines, but not all, because the mechanism inside the cells are different.
Or you can try not using FBS when u want to incubate the cells with the extract, starvation allows the cells uptake more extracts. :)
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