Hi RG community,
we started cultivation of Hek293 cells in DMEM GlutaMAX from Gibco plus 10 % FCS and Pen Strep. The cells grow pretty slowly presumably due to the medium composition. We´re planning to use them for ordinary Plasmid DNA and/or microRNA transfection. My question is if you know a better medium than the one mentioned above (+/- the supplementation) which is purchasable and would require as little as possible Supplementation (so, with added glutamine). If you use a medium which works fine please let me know. I am looking Forward to receiving your answers.
Best wishes,
Stefan
The medium is quite a good one. COnsider to add something else. In one of our last papers we indicate:
HEK-293T cells were grown in DMEM (Gibco, Thermo Fisher Scientific, Halethorpe, MD) supplemented with 2 mM L-glutamine, 100 μg/mL sodium pyruvate, 100 units/mL penicillin/streptomycin, minimum Eagle’s medium non- essential amino acid solution (1/100), and 5% (v/v) heat-inactivated fetal bovine serum (FBS) (all supplements were from Invitrogen, Paisley, Scotland, UK).
I may give you more details if needed as we use them and they grow really "good"We transfect them perfectly well (also with siRNA)
Are the source of HEK cells a reliable one?
Best
Hi there,
As already mentioned, the media you are using is also quite good. We use the same media composition and never faced problem. Like Franco said, you might want to check the source for possible contaminants and passage number. May be it has got nothing to do with the media but your cells are intrinsically sick, so they are growing slow. You can get a fresh vial from some other source and try to grow them in the same media.
Hope it helps!
I am also agree with above two recommendation but I also want to add one thing that after confirmation of your cell source if it does not shows any problem use the same medium with high glucose concentration and FBS from reliable company. As I faced the problem that when I used FBS from different companies and that results in less adhesion of cells to bottom and poor growth with lowest transfection efficiency, while as I changed FBS from others local companies to Gibco, the cells restored to normal condition with very fast growth having highest transfection efficiencies.
Good Luck.
Hi Stefan,
I also agree with the above suggestions. Having your cell line authenticated via STR profiling would be a good step to ensure they are truly HEK293. Secondly, passage number may be an issue here; if the cells are late passage, they may grow more slowly. Third, you may wish to check for mycoplasma contamination, which is undetectable by eye, and which could also harm their growth. Lonza makes a good MycoAlert kit to test for that.
That being said, our lab grows this cell line in Gibco DMEM or Gibco DMEM/F-12 with no supplements other than Heat-Inactivated FBS from Sigma, with great success.
Hope that helps!
Hi Rafael Franco, the "minimum Eagle’s medium non- essential amino acid solution" is in addition to the DMEM?
Thanks,
Dee
Using the same DMEM/F-12, GlutaMAX + FBS 10%, and I observed the HEK 293 cells are growing slowly. Usually, I use without GlutaMAx, morphology seems a little different with GlutaMax, my cells are still as spheres after splitting even the next day.
Hi Janett I am interested in your answer but I am not sure that I understand it. Can you detail it more? What makes cells grow slowly in your "hands" and what means that are "still spheres" . Thanks
- Badges
- Science topic
- Similar topics
- Geoscience
- Cartography
- Mapping