Synthechol™ Ynthetic Cholesterol for Cholesterol Dependent Cell Culture — Development of Non-Animal Derived Chemically Defined NS0 Medium

Abstract

Traditional serum supplemented cell culture medium is fast becoming outdated as new technology permits the omission of most animal derived protein from the culture system. Cell lines derived from the NS0 myeloma cell line are rising in popularity for biopharmaceutical production, due to their high cell growth potential and their subsequent high production yields. However, because of their cholesterol auxotrophic nature, NS0 derived cell lines are particularly challenging for medium development. The first hurdle lies in the fact that there is no source of non-animal derived cholesterol. Second, cholesterol is not water-soluble and thus requires supplementation at the point of use. To address these issues, Sigma-Aldrich has produced SyntheChol™ (Sigma C1231); a synthetic, non-animal derived cholesterol. Initial results demonstrate that SyntheChol behaves similarly to animal-source cholesterol when dissolved in ethanol and supplemented in Hybridoma Medium, Animal Component Free (Sigma H4409). Furthermore, we have developed a sterile, liquid (500X), ready to add SyntheChol NS0 Supplement (Sigma S5442) with increased stability in cooler conditions. The results showed that SyntheChol NS0 Supplement is a stable and effective production-enhancing supplement as compared to both competitor supplements and natural cholesterol dissolved in ethanol. SyntheChol NS0 Supplement can be added to any medium prior to use. Finally, the development of the synthetic cholesterol, SyntheChol, has offered an opportunity of manufacturing an animal component-free, chemically defined NS0 medium.