I request only those researchers who are familiar with culturing of HEK 293 cells to answer this question. In your experience, which FBS is best suited for trouble free culturing of HEK 293 cells? Please share the catalog number of the FBS which you are using.
Hi Srinivas, there is no specific FBS for the cell line you have to check different list to find the one best working for your cells. The Lots differe in between.
Best Michael
Ask your local suppliers for free test samples, then reserve a larger batch of the sample that suits your cells best. That ensures that a whole project is run with the same serum, which avoids that experiments suddenly give weird results, typically shortly before you start writng up the paper...
Dear Jan-Michael
I agree there is no specific FBS for the cell line. Which FBS are you using for HEK293?
Thanks
Hi we are using FBS from Gibco, life technologies. But again you have to test different batches from different suppliers to find the best for your cells because they all behave a little different and then reserve the batch best working as suggested from Berlinda. Best Michael
we are using FBS 10270 from Gibco..but it can be used for other cell lines too..function will be the same..i have attached some of the information about FBS and its inactivation in case you need.
Hi! We are also using FBS from Gibco. Usually works quite well with Hek 293 cells, but I totally agree with Jan-Michael. You have to test the lot previously.
Best
Alexandra
Use any FBS you can get, HEK293 is not a complicated cell line to culture. If you want to do a lot of cell culture, think about buying 500ml bottles of FBS and aliquot them under sterile conditions in 50ml tubes that you store at -20. The big bottles are up to 30% less expensive (calculated per media bottle) than smaller batches.
I agree that you need to check different lots of FBS with your cell line to see what works best with your cells and your system.
I have some additional advise about FBS. When you perform certain experiments that require serum free media, such as transfection of cells, be sure to use a control - that has the serum free media - along with your experimental group. There are serum response elements in many genes that are differentially expressed in response to factors in the FBS.
I once cloned a gene and performed a large experiment transfecting a cell line with the gene, only to find that the expression level was the same in the non-transfected control cells. Several experiments later, I was able to show that the gene was upregulated in the control cells by the removal of the FBS from the serum - which was required for the transfection process.
Good luck with your cells.
Why not go serum-free? Avoids batch-to-batch differences, no dubious origin of the serum, etc. E.g. see: EX-CELL® 293 Serum-Free Medium (Sigma-Aldrich), Pro293™ CD Serum-free Medium from Lonza, Or adapt your cells: "Adaptation of Recombinant HEK-293 Cells to Growth in Serum Free Suspension", R. Mcallister, C. Schofield, G. Pettman, C. Mannix, Animal Cell Technology: Products from Cells, Cells as Products
2002, pp 367-369
More information on problems with FBS read:
van der Valk, et al, The humane collection of fetal bovine serum ......., Toxicology in Vitro 18 (2004), 1-12
van der Valk et al, Optimisation of chemically defined culture media......, Toxicology in Vitro 24 (2010) 1053-1063
Gstraunthaler et al, A Severe Case of Fraudulent Blending of Fetal Bovine
Serum......, ATLA 42, 207-209, 2014
Article The humane collection of fetal bovine serum and possibilitie...
Article Optimization of chemically defined cell culture media-Replac...
Article A Severe Case of Fraudulent Blending of Fetal Bovine Serum S...
I agree with J.B.F. Van der Valk , I would go serum-free to ensure batch-to-batch consistency. In our extensive testing of 6 different brands specifically for the HEK293 cell line, we found Fetalgro BGS ( https://bit.ly/2kik1pS ) to outperform traditional FBS at a significantly lower cost. Hope this helps!
Other fcs-free media for HEK293 cells (and other cells) can be found on https://fcs-free.org/
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