Hi! For HEK293 cell line growth, recommended media is with or without HEPES buffer?
I really don't think it matters, they will grow in just about anything. We grow ours in DMEM + 10% FBS. No HEPES, but HEPES certainly won't hurt.
Hello Clara Radulescu
Most cells require pH conditions in the range 7.2-7.4, and close control of pH by a buffering system is essential for optimum culture conditions.
Regulation of pH is particularly important in culture and is usually achieved by one of two buffering systems:
(i) a natural buffering system where gaseous CO2 balances with the CO3/HCO3 content of the culture medium, and
(ii) chemical buffering using a zwitterion called HEPES.
Cultures using natural bicarbonate/CO2 buffering systems need to be maintained in an atmosphere of 5-10% CO2 in air usually supplied in a CO2 incubator. Bicarbonate/CO2 is low cost, non-toxic and also provides other chemical benefits to the cells.
On the other hand, HEPES has superior buffering capacity in the pH range 7.2-7.4 but is relatively expensive and can be toxic to some cell types at higher concentrations. HEPES buffered cultures do not require a controlled gaseous atmosphere and you can culture the cells outside the CO2 incubator.
So for HEK293 cells, the culture media is either Eagle's Minimum Essential Medium (EMEM) or Dulbecco's Modified Eagle's Medium (DMEM), supplemented with Fetal Bovine Serum (FBS) to a final concentration of 10% and the cells are grown in humidified incubator kept at 37°C with 5% CO2.
In case you do not have a CO2 incubator and you wish to culture HEK293 cells, then you should use the culture media with HEPES. The recommended final working concentration of HEPES in cell culture media should range from 10mM to 25mM. Higher concentration may be toxic to cells under long incubation period.
Best.
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