Cell culture involves growing adherent cells in an artificially controlled environment for various biotechnological applications. Adherent and suspension cell cultures require optimized conditions for both nutritional (substrate or medium) and physicochemical requirements to achieve high product or cell yield. Adherent cell culture involves surface support for cell proliferation while suspension cell culture do not, as suspension cells grow free-floating in the culture medium.
Most isolated cells from animals/humans (except hematopoietic-derived) are adherent in nature. These adherent cells not only represent a formidable production means for viruses or recombinant proteins but also for disease modeling and drug screening in vitro. By contrast, some adherent cell lines were adapted into suspension and do not need surface support for cell growth. Suspension cell lines are usually cultured in flasks that are not surface-treated and require agitation for efficient gas exchange.
Manufacturing vaccines, biotherapeutics, and more involve cell culture (upstream processing) in its early to late stages to produce the desired product. Readily available culture-treated flasks are used for research and development scale in adherent cell culture, while spinner flasks are used for suspension culture. Since adherent scale-up involves an increase in surface area, from simple culture systems like T-flasks and roller bottles, evolved and reliable technologies like packed-bed bioreactors have been introduced. For those that require scale-up for suspension cell culture, traditional stirred-tank bioreactors are used.
Most applications rely heavily on producing high quantities of adherent cells for vaccine production, cell-, and gene therapies. The current large-scale production of adherent cells requires many pieces of roller bottles and cell stacks, among others. These systems pose challenges due to its labor-intensive, time-, and space-consuming factors. With advanced technologies, one can opt-out from scaling out to scaling up.
Put bioprocessing in action with Tide Motion. The adherent Tide Motion technology increases your upstream performance with its automated cell culture and harvesting process, reliable scale-up capabilities, and operational simplicity. The bioreactors follow the Tide Motion principle, which is the gentle upward and downward motion of the culture medium in the vessel. This constitutes an environment with extremely low shear stress, high aeration, and nutrition exchange, where adherent cells can grow well and produce high yield products.
Be at ease whether you are culturing adherent cells for vaccine production, cellular agriculture, gene-, cell therapy, monoclonal antibodies, and more.
Specific culture media and supplements that contain compounds and appropriate source of energy to promote cellular growth
Different seeding configurations in Tide Motion bioreactors
Bioreactors for supporting cell growth of mammalian cell lines such as VERO, MSC, MDCK, HEK293T and more. It provides a in vivo like environment for better cell growth with a higher product yield
Cell surface attachments for anchorage-dependent cells that are macroporous and provide a large surface area for growth and expansion
Cell harvesting systems for faster, closed, and automated way of obtaining cells from the Tide Motion bioreactors
Reagents and monitoring systems for content concentration monitoring and techniques for observing cell growth and development.
Expansion processes for both stem cells and virus on 2-dimensional systems limit its potential to reach high product yield. These systems are enough for research and development scale which only caters to a small number of patients. As these applications evolve, bigger systems are required to achieve high amounts of virus and cells due to how these products are rather different but best when cultured in adherent.
Esco VacciXcell’s TideXcell® Tide Motion bioreactor is designed to produce higher viral titers through its proprietary 100% media exchange and more than 90% stem cell harvest with the TideXcell® Harvesting System (TXLHS). This flexible technology provides cells with 3-dimensional in vivo environment, with extremely low shear stress and high productivity. With scalability of up to 100 L packed-bed volume, one can simply replace 100,000 pieces of flasks to save labor, time, and space requirements. TideXcell™ is available in both single-use or multiple-use technology with advanced, automated controls, equipped with pH, dissolved oxygen (DO) sensors, and sampling ports for cell monitoring and observation. This system is currently used worldwide for vaccine and cell therapy production, helping them bridge the gap of cell culture from discovery to delivery.
White papers for vaccine and viral production on Tide Motion bioreactors tackle the linearly-scalable capability of the technology for culturing adherent cells grown from bench-scale to TideXcell pilot scale bioreactor. This robust cGMP compliant platform allowed the production of higher viral titers and cells to break the barriers in cell manufacturing and cater to many patients.
The term ‘Tide Motion’ is derived from the cyclical high rise and low rise of the bodies of water on earth. In bioreactors, this is the gentle upward and downward motion of the culture media for adherent cell aeration and nutrition exchange. This submerge and emerge principle allows cultured adherent cells to produce quality products that are of high yield.
The cells atteched to the carriers are exposed to air
Determines how long the carriers will be exposed to nutrition
The cells attached to the carriers are exposed to nutrition
Determines how long the carriers will be exposed to aeration
5.5g of carrier is already equivalent up to 15,600 cm2 of surface area(surface area varies depending on cell line morphology)
5.5g of carrier can culture up to 2-3x109 VERO cells and 200 million mesenchymal stem cells (MSCs)
Cells are fixed within proprietary BioNOC II® macrocarriers and are separated from the media. This makes cell harvesting or media harvesting much easier
Dual oxygenation action provides cells with ample oxygen regardless of the bioreactor scale.
Tide Motion bioreactors use separate vessels for culturing and stirring this isolating the cells being cultured from any form of bubbling or foaming.
Freezing cells in Go and G1 phase by nutrient starvation by deprivation of glucose to boost productivity of protein expression.
parameters developed in academic / industrial R&D can be easily translated into clinical trials and commercial production as parameters are identical and hence linearly scalable without complex scale up bioprocess calculations (as those found in Stirred tanks).
Gentle upward and download tide motion (2mm/s) provides nutrients and oxygen to cells without applying shear stress, perfect for shear sensitive cells. Minimizes impurities with host cell protein and DNA/RNA.
Controlled cell growth during cell culture to promote productivity in batch mode via extending exposure time during tide motion without requiring additional feed control
Ideal for biosomilars/biobetters, monoclonal antibodies (MABs), and recombinant proteins. As cells are immobilized, cell retention devices which are typical for perfusion-based suspension cells such as Chinese Hamster Ovary (CHO) are not required to separate cells from media. This means no fouling, foaming, and scalability issues resulting from flow rate limitations of cell retention devices.
5.5 g of carrier (amount of macroporous carriers in 1 CelCradle bottle) is already equivalent to up to 15,000cm2 of surface area. These carriers mimic the cells in vivo state resulting in higher cell or product yield.
Anchorage-dependent cells require surface to adhere in order to grow. Culture vessels such as t-flask and roller bottles are typically used but the difficulty in scale up has led to the development of carriers. Carriers provide optimum environment for adherent cell culture. The advantages of carriers are ease of scale up, cost reduction from serum and culture media, minimum risk of contamination, and reduction of handling process. Carriers are used to grow virus-generating or protein-producing adherent cells in a large-scale commercial production of vaccines and biologics.
Carriers come in different shapes and sizes. Microcarrier beads are spherical in shape and suitable in stirred tank bioreactors. Another form can be in sheets or fibers which are used in packed-bed or tide motion bioreactors. A wide variety of carriers are available in the market with different physical and chemical properties. The choice of carriers depends greatly on the type of cells to be cultured as different cells have varying anchorage requirements.
The BioNOC II® is a macroporous carrier that supports the growth of anchorage-dependent cells including animal, mammalian, and insect cells in either serum-containing or serum-free culture media. It is made of 100% pure polyester nonwoven fabric manufactured according to cGMP guidelines. The special 3-dimensional design and surface treatment on the carrier enhance fluid mixing, immobilization efficiency, protection from shear forces, and nutrient transfer during cell culture. Adherent cells have larger surface area to grow without compromising equipment footprint (Figure 1).
The media reservoir varies from one Tide Motion bioreactor to the other. The research and development scale, CelCradle® system both have the packed bed and the reservoir in the same vessel. For production or manufacturing scale, the TideXcell® separates the media reservoir from the packed bed (Figure 2). This solves scaling up issues regarding aeration exchange. Dual oxygenation is possible through exposing the carriers to air during aeration phase as well as the available dissolved oxygen in the well-mixed culture media (Figure 3). Aside from that, the separation allows dual temperature control, which is an important factor for some cell cultures to achieve high product yield.
Figure 2. A diagram showing the difference in Tide Motion principle for bench-scale system versus manufacturing-scale system.
Figure 3. The separation of the matrix vessel from the mixing vessel solved aeration issues when scaling-up to a larger system.
Tide Motion is a bioprocess method that solves the adherent scale-up problem and provides a single-use bioprocessing method with closed, automated cell harvesting from seed preparation (0.1 L packed bed volume) to production scale (5,000 L packed-bed volume - bioequivalence of 50,000 L in suspension) all in the same 3D Tide Motion (Figure 4).