Lot differences in FBS - (Sep/15/2008 )
Dear forum
Is it still important to consider differences between different lots of FBS or are the methods from the companies (GIBCO) so standardised that this is no longer an issue?
I have been taugth to take batch variation seriously, but have realised that not all labs are as concerned with the matter. Maybe it depends on the biological questions asked?
A trivial question perhaps, but I hope for some feedback
Regards
Kirsten
Department of Haematology
Aalborg Hospital
Aarhus University Hospital
Hi Kirstenf,
Most cell culture labs consider evaluating different FBS lots before ordering a whole lot of it. If you rely a lot on cell culture, then you might want the best for the cells.
I think there is lot variability because there is animal variability, country variability. Also serum has to many proteins to be standardized among each lot.
Is it still important to consider differences between different lots of FBS or are the methods from the companies (GIBCO) so standardised that this is no longer an issue?
I have been taugth to take batch variation seriously, but have realised that not all labs are as concerned with the matter. Maybe it depends on the biological questions asked?
A trivial question perhaps, but I hope for some feedback
Regards
Kirsten
Department of Haematology
Aalborg Hospital
Aarhus University Hospital
Dear Kirsten,
" A trivial question"......IT IS VITAL TO BATCH TEST FBS. The best quality serum is reflected in the cost and availability. New Zealand FBS is the best in the world and the most scarce. If you test viability and grow only, as most companies do, then batch variation is small. HOWEVER the more sensitve the test the greater the variation.
For example, we test:
Cytochrome C concentration in intact cells.
iNOS enzyme induction.
Sodium channel expression in HEK cells.
cGMP in smooth muscle.
Oxiduction/Reduction states of mitochondrial complexes.
Cellular respiration
All the above are affected by FBS batch variation. We buy 50-100 bottles (500ml) of FBS, batch testing at least 3 different batches. We look for the batch which gives us comparable results from the last batch of serum.
The art of experimentation is to reduce as much as possible known variants. In tissue culture these are:
FBS/FCS Batches.
Fyriting Incubators.
Mycoplasma testing all cells.
Monitoring Incubator temperatures.
Hope this is useful
Kindest regards
Rhombus
Is it still important to consider differences between different lots of FBS or are the methods from the companies (GIBCO) so standardised that this is no longer an issue?
I have been taugth to take batch variation seriously, but have realised that not all labs are as concerned with the matter. Maybe it depends on the biological questions asked?
A trivial question perhaps, but I hope for some feedback
Regards
Kirsten
Department of Haematology
Aalborg Hospital
Aarhus University Hospital
there is still some uncertainty between lots since fetal serum as biologic material has some variation; therefore, some companies also launch fully synthetic medium to circumvent variation of medium ingredients; try those if fetal serum is critical...
I totally agree with the Bearer... our lab was totally reliant on cell/tissue culture. We found that we could switch to a synthetic FBS and it did suit some of our cell lines.
Moreover, as it was synthetic it did allay some fears regarding batch variations. Going synthetic maybe worth considering.
Dear Rhombus
Thank you for the input. The procedure you describe with batch testing is exactly how I have been taught to think about serum. So know it's up to me to persuade my new colleagues that this is the way to do it.
Best regards
Kirsten
Is it still important to consider differences between different lots of FBS or are the methods from the companies (GIBCO) so standardised that this is no longer an issue?
I have been taugth to take batch variation seriously, but have realised that not all labs are as concerned with the matter. Maybe it depends on the biological questions asked?
A trivial question perhaps, but I hope for some feedback
Regards
Kirsten
Department of Haematology
Aalborg Hospital
Aarhus University Hospital
Dear Kirsten,
" A trivial question"......IT IS VITAL TO BATCH TEST FBS. The best quality serum is reflected in the cost and availability. New Zealand FBS is the best in the world and the most scarce. If you test viability and grow only, as most companies do, then batch variation is small. HOWEVER the more sensitve the test the greater the variation.
For example, we test:
Cytochrome C concentration in intact cells.
iNOS enzyme induction.
Sodium channel expression in HEK cells.
cGMP in smooth muscle.
Oxiduction/Reduction states of mitochondrial complexes.
Cellular respiration
All the above are affected by FBS batch variation. We buy 50-100 bottles (500ml) of FBS, batch testing at least 3 different batches. We look for the batch which gives us comparable results from the last batch of serum.
The art of experimentation is to reduce as much as possible known variants. In tissue culture these are:
FBS/FCS Batches.
Fyriting Incubators.
Mycoplasma testing all cells.
Monitoring Incubator temperatures.
Hope this is useful
Kindest regards
Rhombus
Hi Kirsten,
yeah it very imp to consider FBS lot variation. I have a worst experience with changng FBS lot. By changing FBS lot (PAN) I just lost response in functional assay in my chemokine receptor expressed recombinant cell line. Then I had to order the same lot of FBS and could get functionality in cells again afet a month.