HEPES is a buffer reagent used for cell culture, with a buffer range of 6.8 to 8.2. In open cell culture conditions, its pH value is often affected by external factors. If a HEPES buffer is added, it can prevent pH oxidation in the culture medium from increasing. Therefore, during cell culture, HEPES buffer is often necessary. In addition, it is also widely used in the uptake of organic molecules, changes in oxidative stress mechanisms, or inhibition of ions.
Compared with bicarbonate, the buffer solution of HEPE has higher solubility and stability, and has the smallest complexation with metal ions. It has been proven that HEPES can be responsible for significant modifications of organic molecule absorption and other parameters in biotechnology under different usage scenarios. There are currently several molecular mechanisms for oxidative stress and ion channel inhibition.
Prion, also known as prion, protein infection factor, toxic prion or infectious protein, is a small molecule non immune hydrophobic protein that can infect animals and replicate in host cells. In the process of seeking prion gene therapy in our laboratory, we infected Lentivirus on cells with prion in the supernatant of HEK 293T containing related proteins. In the control experiment, we observed an abnormal subtype of accumulated prion protein (PrPSc) with significant inhibition by using unrelated Lentivirus or culture medium alone, and finally confirmed that it was related to the existence of HEPE in the culture medium.
When cell culture medium supplemented with HEPES is used on prion infected cells, there is a significant concentration dependent inhibition of accumulation of prion protein abnormal subtype (PrPSc). This effect only exists in living cells and has not been proven to be related to changes in PrP environment or biology. This inhibition of HEPES can also be used to prevent prion contamination or reproduction in cell culture.