HEPES buffer in cell culture

Details:

HEPES, as a part of the buffer, has its traces in many places. Just because we don't know, we often ignore its existence, and forget that it itself has a role we can't imagine in many places. Especially in our cell culture, it has a unique advantage. For you who don't know about cell culture and HEPES buffer, here is a good solution to why cell culture can't be separated from HEPES buffer.

What is cell culture

Cell culture is the clone of cells, which can get a large number of cells or their metabolites through cell culture. Because biological products are all from cells, cell culture technology is the most core and basic technology in biotechnology. But when it comes to cell culture, we have to mention HEPES, a amphoteric buffer.

Why cell culture is needed

This is because the growth and metabolism of cells in culture will lead to drastic changes in the environmental pH of the medium. Therefore, in order to maintain the environment in which the cells can be stably cultured, it is necessary to add some biological buffer to the medium.

Why use HEPES buffer

The best culture environment for most cells is pH 7.2-ph 7.4  ， The buffer range of HEPES is just within pH 6.8-ph 8.2, which is beneficial to cell culture. HEPES has higher stability in maintaining the pH value of cell culture medium compared with other buffer solutions (such as PBS (phosphate buffer salt water) and Tris, which is also the reason why HEPES is widely used in cell culture, tissue culture, protein purification and extraction, immunoprecipitation, cell cleavage, living cell imaging and other biological / biochemical research.

What should HEPES buffer pay attention to in cell culture

1. The main initial step of adding HEPES to the medium is to maintain the cell environment in a stable pH value. This is important, especially when it takes a lot of time to work on cells outside the incubator, because it can lead to pH changes.

2. It is important to note that sodium bicarbonate is another buffer used to stabilize the pH of the medium. However, HEPES buffers should not be used in place of sodium bicarbonate, so that the acidity of the medium will be stronger than expected. Because the normal bicarbonate buffer is CO2 dependent, and HEPES buffer is CO2 dependent. This is why the medium is only placed in the indoor air, not in the incubator, and the phenol red in the medium changes color over time.

3. Here is a question to consider. What type of primary cell is needed in cell culture Some primary cells are very easy to cultivate, and do not need to supplement the medium in large quantities, while others are very picky, and they need to be highly buffered and supplemented.

4. The concentration of HEPES buffer in cell culture medium is recommended to be 10-25mm, and higher concentration may cause toxicity to cells during long incubation.