The Extensive Usage of HEPES in The Field of Cell and Tissue Culture

These cells may be grown in a culture medium of biological origins such as blood serum or tissue extract, in a chemically defined synthetic medium or boosted, and aided by growth supplements such as (FBS) Fetal Bovine Serum. These tissues can also be cultured, or cells can be made to form tissues in the culture, with the express usage of supports like sponge gels that are immersed in various culture mediums. A medium must contain proper proportions of the necessary nutrients for the cells to be studied and must be appropriately acid or alkaline.

These cultures allow for more sophisticated studies like tissue formations, differential cell-cell interactions, tissue-specific gene regulations, and accurate drug-response testing, such as with tumors. In addition, these cells in tissue cultures are subjected to a broad range of experimental treatments—for example, hormones, viruses, vitamins, drugs, disease-causing microorganisms, or suspected cancer-producing chemicals may be an additional element to the culture. Researchers then observe the cells, looking for global changes in cell behavior or function or changes in specific molecules, such as alterations in the expression of specific proteins or genes.

There are many specific chemicals or other reagents that are specifically used for a variety of purposes centered around cell and tissue cultures. One such reagent is HEPES, which is widely known to be used for animal cell and tissue cultures.

HEPES has been described as a particularly effective all-purpose buffer available for biological research. At a specific biological pH, the molecule is zwitterionic and effective as a buffer at a pH range of 6.8–8.2. HEPES has been used in a broad range of applications, including tissue culture. It is most commonly used to buffer cell culture media in air. HEPES buffer does not necessarily confer cytotoxic effects on cells, which makes it become usable in animal cell cultures.

The express growth of animal cells in a nutritionally complete tissue culture medium is usually most optimal when the specific medium is buffered at a particular pH in the range of 7.2–7.4. Therefore, for maximum functional efficiency, the pKa of the chosen buffer should be as close to the required pH as possible.

Slight effects described of HEPES on plaque titrations are very probably because of the more exacting bicarbonate requirement of vaccinia virus as compared with other viruses. The specific yield of cells grown in a HEPES-buffered media—determined by both total and viable counts— was the same as that in unmodified media or even slightly greater with certain lines of cells.

The specific culturing of cells in a laboratory has some very specific purposes. From diagnosing certain types of diseases and genetic profiling to testing the efficacy of antibiotics, identifying microbe species, and even experimenting on cancer cells and extracting molecules, research on cell cultures has come a long way.