We use to use FBS form Gibco to culture our colon cance cell lines (HCT-116, COLO-205, HT-29 and SW-620) and it were o.k, with good morphological appearance as in the database, I sub-cultured them every 2-3 days. But recently we started using FBS from another company, but we realize the cell's growth are very low, we subculture them every 8-10 days, and even the morphological forms have changed. So could change of FBS (Fetal Bovine Serum) affect the quality of the cell?
You may not see the effect of the new FBS at first - cells can have intra-cellular stores they can use for their initial growth (this is one of the reasons why I test for ~ 2 weeks - to allow the FBS they had grown in previously to wear off although to be honest this should disappear within a few days).
You may not even see an effect at all unless the cells are stressed (such as plating at low density or plating in the presence of toxins etc). When stressed the cell may rely upon factors in the FBS (such as growth factors) to keep alive/dividing which may be absent/lower in the "new" FBS
.
When I FBS test I deliberately grow the cells at the lowest "split" level recommended (i.e. if the cell product sheet says inoculate new cultures between 1x10e5-3x10e5 I will split at the lowest level - this is the second reason I grow for up to 2 weeks, re-setting the cell densities in all of the cultures each time I split). This helps highlight differences between FBS batches.
Also some FBS types will be better for certain cells. You may find FBS from "supplier X" supports the growth of Jurkat cells better than FBS from "supplier Y" but your HeLa cells might grow better in the FBS from "supplier Y" than they do in FBS from "supplier" X (if that makes sense?). In fact I noticed this effect when testing Gibco and ATCC FBS with those lines that I mentioned above (although it was a small effect and in general good quality FBS can support most cell types well).
If you can't go back to your old FBS supplier then you can try to "serum adapt" your cells by cultivating them for a time in your new FBS - if they don't die they may eventually increase in growth rate/viability as the cells adapt to their new conditions.
However this "selective pressure" may cause the cells to lose or gain some phenotypic functions (all cells in culture are under a degree of selective pressure which is why we make working banks of cells and do not over-passage them as they will eventually change (phenotypic drift)).
Phew! I hope all that makes sense!
Gary
Hi Bashir - this is a big YES.
FBS varies in quality company to company. If you are changing your FBS supplier you must perform batch testing (you grow the cells in your "new" FBS alongside your "old" FBS and note d.t., viability etc). You need to keep the cells growing for at least 2 weeks (with splitting) in the different FBS to make sure the new FBS is not having an effect. Note: Cells can become adapted to a certain media in culture so it's not impossible if you keep passaging the cells in the "cheaper" FBS sources they may get used to it (but for consistency between old and new experiments this is NOT recommended).
I recently tested 3 lines (Jurkat, AU565, MCF7) with FBS form 4 different suppliers.
The "cheaper" FBS (suppliers X & Y) were a lot worse than FBS from ATCC or Gibco (who supply high quality FBS).
The FBS quality can be affected by the health of the animals it is from - if the animals have been stressed, ill or treated with antibiotics this can all potentially carry over into you cell culture media.
IMHO buying cheap FBS can be a false economy in the long run.
I hope this helps,
Gary
Yes .Definitly the FBS origin matters in the maintainance of cells. Use Australian origin FBS from a standard company like Gibco.
Thanks Gary and Reddy, for the nice comments, and it are v useful.
So if I want to start thaw a new batch of my cells from liquid nitrogen, that I already stored using Gibco FBS, would the effect of the new FBS persist? or the effect will be less?
You may not see the effect of the new FBS at first - cells can have intra-cellular stores they can use for their initial growth (this is one of the reasons why I test for ~ 2 weeks - to allow the FBS they had grown in previously to wear off although to be honest this should disappear within a few days).
You may not even see an effect at all unless the cells are stressed (such as plating at low density or plating in the presence of toxins etc). When stressed the cell may rely upon factors in the FBS (such as growth factors) to keep alive/dividing which may be absent/lower in the "new" FBS
.
When I FBS test I deliberately grow the cells at the lowest "split" level recommended (i.e. if the cell product sheet says inoculate new cultures between 1x10e5-3x10e5 I will split at the lowest level - this is the second reason I grow for up to 2 weeks, re-setting the cell densities in all of the cultures each time I split). This helps highlight differences between FBS batches.
Also some FBS types will be better for certain cells. You may find FBS from "supplier X" supports the growth of Jurkat cells better than FBS from "supplier Y" but your HeLa cells might grow better in the FBS from "supplier Y" than they do in FBS from "supplier" X (if that makes sense?). In fact I noticed this effect when testing Gibco and ATCC FBS with those lines that I mentioned above (although it was a small effect and in general good quality FBS can support most cell types well).
If you can't go back to your old FBS supplier then you can try to "serum adapt" your cells by cultivating them for a time in your new FBS - if they don't die they may eventually increase in growth rate/viability as the cells adapt to their new conditions.
However this "selective pressure" may cause the cells to lose or gain some phenotypic functions (all cells in culture are under a degree of selective pressure which is why we make working banks of cells and do not over-passage them as they will eventually change (phenotypic drift)).
Phew! I hope all that makes sense!
Gary
Hi Bashir,
We noticed in our lab that using the same FBS but from a different Lot might affect your cell growth (we use Gibco, not a cheap one), therefore we usually reserve a whole batch, and when this Lot is over, we perform quality tests on the new batch.
Some companies provide tissue culture tested ones, which are of course more expensive.
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