Variability in human fibroblasts normal?

[sciencegal]

Please help...I've been working with human fibroblasts for some time now and am finding it nearly impossible to replicate any results as the same cell line does not yield consistent results over time. We have frozen stocks ranging anywhere from about passage 5 through 15 and do not have access to "fresh" supplies so I am forced to passage over time and periodically (about every 2-3mos), I will go back to my frozen stock and thaw a new vial so I don't have the same cells in culture for more than 3 months. I don't know what to do and neither does anyone else I work with and I'm not sure how much longer I can pursue this. I have tons of data but none of it is very useful b/c my results are constantly changing and with no one to guide me, I am very close to just giving up. Any advice/experience would be greatly appreciated!

[Anastasia_S]

I work with human macrophages and although I isolate them from blood rather than passage, I find quite a lot of variability from batch to batch - the donors are different and also, it seems that as they grow, some responses change
Try blocking your data by the "age" of your fibroblasts
also, slight (and not!) variability with such cells is very common

[aksoyi]

Hi sciengal,
i'm sorry I can not really help you, i think it's normal to have variability but as anastacia said you should do yiur experiment always at the same passage with your different cells.
Actually, I have a question for you, where did you get all your fibroblasts samples ? I'm looking for fibroblasts to do some genotyping on them and I need at least fibroblasts from 10-15 different patients. Could you help me for thsi pease ? Thanks a lot,

Irene