I'm new to primary human fibroblast culture and I'm studying aging using cells from old and young donors. Some research groups have used media supplemented with FGF2, so I cultured cells in the presence and in the absence of it. Cultures without FGF2, when they become confluent, they immediately change morphology to become 'flatter'. Does this mean they're senescent? However doubling rate is very high (maybe too high for a 60year old donor) for both cultures. I was hoping that without FGF2 the cells would proliferate slower to allow longer treatments with drugs etc. Does the morphology of either culture look better? Any feedback is much appreciated!
Hello Elena Pa,
Culture fibroblasts with FGF-2 because cells grown on plastic are not nearly as happy as they are in the body. So, feeding cells with FGF-2 helps signal them to proliferate in order to grow more.
FGF-2 is known to allow fibroblasts to maintain their phenotype more effectively. Cultures without FGF-2 results in cell growth decay with an aberrant phenotype (flattened morphology).
Whenever you use cells for any cell-based assay, try not to allow cells to deviate from their original characteristics, otherwise you may end up interpreting the results wrongly.
From the image, I can observe the difference between the two groups, with +FGF-2 group looking far much better at day 3.
Best.
Dear Elena Pa
If you want to check the effect of FGF-2 on cell growth, you should preferably use the alpha MEM culture medium with as little FBS as possible.
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