What is the performance of HEPES buffer in cell culture

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Cell culture is often carried out in biomedical laboratories. The requirements for physiological environment during cell culture are relatively high. In the whole culture system, pH value plays a vital role in the success or failure of the experiment. HEPES (4 - (2-hydroxyethyl) - 1-piperazine ethanesulfonic acid) is an amphoteric organic chemical buffer suitable for most biological pH ranges. It is classified as a general buffer that can be used in biological research.

HEPES buffer

Therefore, HEPES is often used as a buffer in cell culture medium. Compared with the bicarbonate / CO2 buffer system, it works better to ensure a stable pH value during the treatment of cell culture medium outside the incubator. Due to the inherent charge of HEPES, it is considered that it will not be absorbed by cells, which is also a prerequisite for good and his colleagues in the development of cell culture buffer.

In the past few years, some data have shown that HEPES seems to be absorbed into cells in cell culture and will affect cell function. Therefore, the performance of three different cell lines (MCF-7, U2OS and HeLa) in HEPES containing medium was studied. The results showed that they all adapted to the medium containing HEPES. They survived and proliferated in the presence of HEPES. The determination of intracellular metabolites showed that HEPES existed in all cell lines. Further analysis of MCF-7 cells showed that HEPES in cells could be detected even 48 hours after changing from HEPES containing medium to HEPES free medium. This effect may be improved if the concentration of HEPES is reduced during use.

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