Abstract
Recent cell culture media for mammalian cells can be abundantly formulated with nutrients supporting production, but such media can be limited to use in host cell culture, transfection, cell cloning, and cell growth under the low cell density conditions. In many cases, appropriate platform media are used for cell line development, and then replaced with rich media for production. In this study, we demonstrate rich chemically defined media for Chinese hamster ovary (CHO) cells that are suitable as basal media both for cell line development and for final production of culture process. Set up for transfection, semi-solid media optimization, mini-pool screening, and single cell cloning media development were performed, and final clones were obtained with higher productivity in fed-batch culture mode using rich formulated media comparing with lean formulated media. Developed methods may remove the requirements for cell adaptation to production media after cell line development, and relieve the clonality issues associated with changing the culture media. Furthermore, established methods have advantages over traditional approaches, including saving resources and decreasing the time and the effort required to optimize the production process.
Highlights
The cell culture media are necessary to supply sufficient nutrients for the cell growth for mammalian cells such as Chinese hamster ovary (CHO) cells, and applying appropriate media is important in developing cell line and cell culture process with high productivity and quality
CDM2, CDM3, and CD FortiCHO (Thermo Fisher Scientific, Waltham, MA, US) worked well in our monoclonal antibody cell line and recombinant protein cell line which are developed from our cell line development (CLD) platform using CDM1
We attempted to adapt CHODG44 cells already adapted to CDM1 to these media, and cells only failed to adapt to CD FortiCHO
Summary
The cell culture media are necessary to supply sufficient nutrients for the cell growth for mammalian cells such as Chinese hamster ovary (CHO) cells, and applying appropriate media is important in developing cell line and cell culture process with high productivity and quality. After SFM was available, the cell line was adapted to SFM to boost up the productivity, apply to suspension cell culture process and relieve the issues using serum (Fig. 1a). Current CLD commonly simplifies the process by employing SFM-adapted host cells and developing the cells in serum-free conditions in order to reduce the time of adapting cells in SFM at the later stage (Fig. 1b). Technically, this requires optimization of the host cell media, transfection media, selection media, and the cloning media used in each step of CLD. Setting up a new CLD platform with SFM consumes the significant resources to optimize the media in each step and may use the different media from the basal platform media in a particular step
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