Serum-free adaptation (serum free media, cell suspension culture) is the process of growing mammalian cells in specialized media that replaces serum (typically fetal bovine serum) to meet FDA Standards for biologics production.
Mammalian cell culture allows a manufacturer to produce high quality biologic products. However, optimization and scale up for production present unique challenges. Slow growth rates and inconsistent productivity are limiting factors in large-scale mammalian cell culturing processes. Current technologies have answered some of these challenges through the use of highly efficient bioreactors.
We offer the following services around cell adaptation:
- Direct adaptation to serum-free media
- Sequential serum-free adaptation
- Selective cell suspension culturing for bioreactors
- Adaptive cell suspension for bioreactors
Contact one of our service managers with specific questions about serum-free or suspension adaptation.
Below you will find some more detailed information about our Cell Serum-Free Adaptation services.
Large scale-up of mammalian culture systems means more cells are involved. More cells means that there will be an additional depletion of nutrients and oxygen within a loosely defined cellular medium, and a build-up of toxic metabolic by-products. Culture media is optimized to support cell growth, but most media contains an undefined serum portion (5-20%). The FDA discourages the use of serum containing media for biologic product production because of potential pyrogens and contaminants including mycoplasma, viruses, and fungi.
BioReliance specializes in serum-free adaptation of cell culture systems for therapeutic scale-up and production. Standardization of media to be serum free improves reproducibility and decreases contamination.
Serum free adaptation protocols
Serum free adaptation of mammalian cell culture can be performed in two ways; Direct Adaptation or Sequential Adaptation (weaning).
Direct Adaptation is performed with a cell line that is at 90% confluent grown in normal media that is directly sub-cultured into serum-free media optimized for the particular nutrient needs of a particular cell type. This method is usually sufficient for transformed cell lines that have a simpler overall nutritional requirement.
Sequential Adaptation (weaning) involves decreasing the concentration of serum found in the media over a defined number of passages, until the cells are adjusted to media lacking serum.
For example, passage 1 will have 75% media with serum and 25% serum-free media. Passage 2 will have 50% media with serum and 50% serum-free media, and so on until the desired serum concentration is achieved. Some cells may need to be passaged two or three times at a particular stage to adjust to the reduced amount of serum.
BioReliance has vast experience in both methods of serum free adaptation. Contact one of our service managers with specific questions.
Adherent cells present difficulties in scale-up. Many bioreactors and cell factories require cell suspension cultures for proper growth and biologic production. BioReliance has protocols to persuade adherent cells to grow in suspension.
These methods include selection and adaptation. The selection procedure involves disturbing a confluent cell layer and selecting the cells that are able to grow loosely attached to the monolayer through centrifugation. These cells are then cultured in suspension and selected for production of a therapeutic of interest. The adaptation method on the other hand, involves taking the cells from the selection method and applying varying degrees of agitation to maintain suspension culture.
Contact a BioReliance research professional about specific cell line adaptation needs or difficult to adapt cell lines.