Development of DMSO-free Cryopreservation Media for Cells Producing Biopharmaceuticals

다국어 초록 (Multilingual Abstract)

Many studies have shown that cells are likely to experience genetic or phenotypic variations over generations. The only way to prevent this is to keep the cells frozen at extremely low temperatures ( -80℃ to -196℃). However, this cryopreservation method can have a negative effect on cells because it provides cells with an extreme environment (high osmotic pressure, physical and chemical stress, etc.). To reduce this negative effect as much as possible, CPAs (cryoprotective agents) are used. The most representative CPA is DMSO (dimethyl sulfoxide). However, the use of DMSO is somewhat controversial as studies have shown that it may reduce cell viability or affect gene expression systems. Therefore, we conducted research to develop preservatives using substitute substances other than DMSO. Similar substances were evaluated to select two CPAs as highly permeable substances such as DMSO were considered key candidates. These can prevent ice formation and inhibit physical damage to cells by removing water from the cells. However, recently, the freeze protection effect of antioxidants was confirmed as it has been argued that not only the physical protection of cells but also the chemical damage to ROS outbreaks should be controlled. In conclusion, we identified cell-permeable substances that have the key effects to replace DMSO, and additionally designed cryopreservation compositions by adding functional additives. Like this, we aim to eliminate controversial DMSO when freeze-preserving cells for biopharmaceutical production, and to develop products with equal or better freeze-preservation capabilities without using DMSO.