Cultured cells are excellent hosts for the propagation of many types of viruses. The ability of cell culture systems to produce large quantities of attenuated viral particles has served as the basis for the production of both human and veterinary vaccines. Traditional methods have relied on the production of viral agents in cells cultured in medium supplemented with serum, most commonly fetal bovine serum (FBS). The animal serum in cell cultures can cause a number of problems. The increased costs of raw materials and post-production processing associated with serum have prompted interest in the development of serum-free media for vaccine production. More recently, the potential for contamination by adventitious agents present in serum has heightened regulatory concerns regarding the use of animal-derived components in media used for pharmaceutical manufacturing. Therefore, we have developed media for vaccine production with reduced levels or completely devoid of animal-derived components. We will also continue our commitment to traditional manufacturing methods by maintaining and expanding the range of basal media for use with serum supplementation and serve as the basis for customization into serum-free formulations.
The ability to genetically engineer viral particles for use as therapeutic agents to treat genetic diseases (i.e., gene therapy) represents an exciting avenue of science and technology. The regulatory concerns regarding the use of animal-derived materials in media used in pharmaceutical manufacturing, particularly for therapeutic injectables, have led to recommendations that all animal-derived components be avoided when therapeutic agents are manufactured. We offer media tailored to the needs of the two most common cell lines used for the propagation of viral particles used in gene therapy. These media are formulated without the use of animal-derived components.
EX-CELL® Media is an animal-protein free, serum-free medium developed for long-term cell growth. The cells, in an attachment culture, can be subcultured directly into EX-CELL® Media from serum-supplemented media without adaptation. Cell densities and doubling times achieved under serum-free conditions are comparable to those achieved in a serum-supplemented culture.
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Human and veterinary vaccines have typically been manufactured using chicken eggs and primary chicken embryo fibroblasts (CEFs) (Table 1). These cell substrates pose a number of limitations including risk of insufficient supply, time-consuming processes with inconsistent yields, high costs of manufacture, the potential for allergic responses to egg-components and concerns associated with using bovine sera in CEF cultures.
A fully documented stable duck cell line, termed EB66, derived from embryonic stem cells has been established as a superior alternative for the cost-effective manufacturing of vaccines currently produced in CEFs or eggs. EB66 cells maintain genetic stability, are immortal and grow in serum-free media. An added advantage is high-density growth as suspension cells without the need for microcarriers.
The EX-CELL® EBx™ media and supplements are optimized for use with suspension EB66 cells (Valneva SE) to ensure maximum growth and high titer viral vaccine production. High cell viability and high cell density growth in suspension provides the optimal environment for high-titer viral vaccine production. Typical titer values range from 8 to 9 log TCID50/mL, when using EB66 cells grown in EX-CELL® EBx™ medium and infected with modified vaccinia virus Ankara (MVA) in stirred-tank bioreactors.
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For EX-CELL® EBx™ media support and information, please contact Technical Service. For EB66 cell line and viral production process development information, please contact Valneva SE.
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Traditionally, viruses for seasonal influenza vaccines have been grown in eggs. Recently, influenza vaccine development has used cell-culture techniques for growing virus in living cells instead of the time-consuming egg-based process.
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Fetal bovine serum (FBS) is the most widely used growth supplement for cell culture media because of its high content of embryonic growth promoting factors. When used at appropriate concentrations it supplies many defined and components that have been shown to satisfy specific metabolic requirements for in vitro cell culture.
We continue our commitment to life science researchers with over 30 years of fetal bovine serum production and manufacturing with extensive product testing (Table 2-3). We offer a variety of origins with special treatments and specifications along with a dependable, consistent supply that is ISO certified quality from collection to finished product.
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