(T0930-09-56) Development of a High Purity, High Productivity Cell Culture Media Additive

Purpose: Mammalian cell culture processes are used across the biopharmaceutical industry as vehicles for the production of proteins and viruses and as therapeutics themselves. Significant research goes into the optimization of cell culture methods to maximize growth, health, and productivity of different cell types. Cell culture media are highly formulated as part of this optimization, with formulators pursuing xeno-free and chemically-defined ingredients.The purpose of this study was to identify methods for purification of methylcellulose to be suitable for use in cell culture and to demonstrate utility of ultrapure methylcellulose in several cell culture processes.

Methods: Cell lines were maintained in humidified incubators according to suppliers’ recommendations. All medias were commercially available from major suppliers and chosen based on to what cells had been acclimated. Adherent cells were plated on TCPS well plates or T-25 flasks. Suspension cells were grown in 125 mL flasks with final volumes of 30 mL or 250 mL flasks with final volumes of 60 mL using a fed-batch processes. Cells counts and viability were measured by hemocytometry and use of WST assays.

Results: Several proprietary process improvements were employed to reduce insoluble impurities by 60%, easing solution sterilization. Improvements also allowed minimization of trace metals and organics, leading to a more chemically-defined product. Application studies show utility of the purified methylcellulose to increase rates of cell growth in several relevant cell culture models. CHO cell peak viable cell density in suspension improved 2-fold with the inclusion of ultrapure methylcellulose in shake flasks. Significant improvement was also shown compared to addition of poloxamer to the media. In adherent cultures, Vero and hMSC cells grew 20% faster with methylcellulose present in the media compared to control.

Conclusion: New, chemically-defined media additives are required to continue to drive process intensification and media productivity. In this work, we have shown the ability to develop a highly purified methylcellulose suitable for use in mammalian cell cultures. >In several application studies, addition of the ultrapure methylcellulose to media led to significant improvements in cell growth rates, allowing for higher cell densities or shorter cycle times for pharmaceutical processes.

Comparison of CHO cell suspension growth in shake flasks with addition of methylcellulose and poloxamer 188.

Addition of methylcellulose improves peak viable cell density for CHO cells in suspension culture in a scaled-down bioreactor.