Troubleshooting Cell Culture Media for Bioprocessing

Sponsored by: GE Healthcare
Session ends: February 21st, 2014, 3:00pm MST
Answers by: Peggy Lio, Director of Process Science, GE Healthcare


Cell culture media, including basal media and feeds, are key elements impacting the performance of bioprocesses. Advancements in cellular and metabolic understanding, coupled with high throughput applications, have led to evolved approaches in medium development and optimization resulting in innovative cell culture media with desired characteristics to meet specific needs.

However, challenges associated with the use of cell culture media still exist.  Examples include manufacturability, subpar stability, inconsistent performance and/or underperformance in terms of productivity and quality attributes.  In some cases, troubleshooting efforts can be guided by learned know-how but in other circumstances a systematic approach is necessary to identify the root cause.  In this Ask the Expert session, we invite you to participate in discussions on experience and lessons learned around cell culture media troubleshooting. Please submit your questions about cell culture media for bioprocessing.

ask the expert

Questions & Answers

What techniques can you suggest for improving media solubility for media that is difficult to get into solution?

There are approaches, such as alternating pH (either acidic or basic) and temperature (mostly warm), that can accelerate/facilitate solubilizing media compounds. However, they don’t necessarily improve solubility (maximum concentration in a saturated solution)issues. Therefore, it is important to also evaluate the composition of your media formulation to assess whether there are components that are at […]» Read More

Can you explain how cell metabolism characteristics can be used to optimize media in CHO cells?

There are several ways of utilizing cell metabolism characteristics for medium optimization. Firstly, the metabolism rate of key medium components such as amino acids and vitamins can easily be determined by spent media analysis. The metabolic profiles can then be used for design of stoichiometrically balanced medium formulations. Since metabolic characteristics are dynamic, utilization of […]» Read More

We are seeing a rise in ammonia in one of our CHO cultures prior to harvest. It isn’t entirely predictable so we are having trouble troubleshooting. Any suggestions?

The concentration of glutamine and/or asparagine in your cell culture media should be assessed and limited since both amino acids can be deaminated leading to ammonia as the byproduct. During late stationary phase of mammalian cell culture, cells may utilize alanine to generate pyruvate and this process also produces ammonia. Supplementation of pyruvate into the […]» Read More

Which serum free media is suitable for optimization of CHO and BHK cells (Cell Culture) producing recombinant blood clotting factors.

There are commercially available serum free media suitable for the optimization of recombinant blood factor expression in CHO or BHK cells. Depending on the cell type and specific molecule of interest, the media may need to be supplemented with other components (e.g., growth factors, cell protection agents, and product dependent factors). Media optimization should always […]» Read More

Is it always important to supplement media with depleted components?

Nutrient depletion causes cell apoptosis which can impact productivity levels and/or product quality therefore it is important to supplement media with depleted components. In order to achieve the best performance, a metabolically balanced feed formulation and an optimal feeding schedule should be designed and applied.» Read More

What media components can reduce apoptosis and increase cell viability beyond critical density?

Apoptosis is typically caused by nutrient depletion and/or metabolite accumulation. Nutrient depletion can be prevented by proper feeding strategies. Metabolite accumulation is less of concern in a perfusion process but is often challenging to tackle in a fed-batch process. Media optimization approaches to control the accumulation of metabolites may include: reducing glutamine to control ammonia, […]» Read More

Are there media with carbon source different from glucose?

Yes, there are media with alternate carbon sources. For example, galactose, which has a lower metabolizing rate than glucose, may be supplemented into cell culture media to remedy the issue of high lactate generation in certain cell culture situations. Disaccharides are also commonly used in culture media for non-mammalian cells.» Read More