One-Stop Solution for Virus Production with 3D Dissolvable Porous Microcarriers
In recent years, there is an increasing number of research and development projects in the fields of oncolytic viruses, viral vectors, and novel vaccines. Many of these emerging viral biological products rely on adherent cell culture for virus production. Under the traditional cell culture process, adherent cells are usually cultured in cell culture flasks, roller bottles, or cell factories. However, these approaches pose a limit in scaling and it is difficult to precisely control the culture process. Moreover, it is hard to ensure consistent cell quality while achieving large-scale cell expansion. Using a microcarrier in combination with a bioreactor for the suspension culture of adherent cells would suggest a better solution.
- Traditional microcarriers are usually rigid solid micro-spheres made of synthetic polymers. The adherent cells grow in a 2D curve over the surface of the microcarrier.
- At a high feeding density, the bioreactor's stirring motion may result in lower cell viability and generation of microcarrier fragments, which can lead to non-negligible consequences such as an increase in cost, safety risk, and lower harvest rate in the subsequent purification stage of virus production.
CytoNiche launched the first 3D dissolvable porous microcarrier - 3D TableTrix® Microcarriers (V Series) for clients in the sector of viral biological products. The V series can be used to culture a variety of adherent cells and demonstrate advantages in cell expansion and virus production compared to the traditional microcarrier.
- Honeycomb porous structure with diameter 30–50 μm and porosity > 90%;
- Customizable with a range of particle size 50-500 μm;
- Effective adherence area >9000cm2/g, about 2-3 times that of a traditional microcarrier, providing more growth capacity to hold adherent cells with the same feeding density;
- Bionic chemical properties featured, which can effectively buffer fluid shear force in a stirred tank bioreactor and mitigate the shear force. Hence, providing a higher cell viability and increasing the viral production yield;
- The microcarrier is 100% degradable. A microcarrier degradation residue detection method (kit) verified by the National Institutes for Food and Drug Control is available.
3D TableTrix® Microcarriers (V Series)
This product supports SIP online sterilization, and customized pre-sterilization packages are also available.
Green, fluorescentspotsrepresent living cells,this figure showsthe growth condition ofVero cells on the microcarrierafter72 hours
Multiplehomogeneous microcarrier particles were observedthroughscanning electron microscope
- The two pharmaceutical excipient products developed by CytoNiche for cell therapy - Gelatin Microcarriers and Gelatin Microcarrier Tablets (Cell) - have received the quality and safety evaluations by the National Institutes for Food and Drug Control (CDE) and have obtained 2 qualifications for pharmaceutical excipients from the National Medical Products Administration (CDE approval registration number: F20210000003, F20210000496).
- The latter (Gelatin Microcarrier Tablet for Cells*) has also obtained the Drug Master Files (DMF) qualification for pharmaceutical excipients from Food and Drug Association (FDA DMF: 35481).
* Gelatin Microcarrier Tablet for Cells*: Spherical microcarrier are preformulated and packaged as tablets.
- The microcarrier GMP certified manufacturing platform also obtained ISO9001 and ISO13485 certifications.
To achieve high quality and large-scale 3D cell culture and after considering hydrodynamics, process control parameters, and other factors, CytoNiche independently developed and launched 3D FloTrix® vivaSPIN Automated Bioreactor to complement its core product and technology - 3D TableTrix® Microcarrier and 3D FloTrix® dynamic culture process.
3D FloTrix® vivaSPIN Automated Bioreactor has a tank designed with top hygiene grades that can achieve reliable closed cell culture in a C + A level environment.
The controller is equipped with the PECALS™ control system which can accurately execute automation program, support remote control of parameters and record detailed real-time information. This allows you to monitor the cell culture process at home.
The PECALS™ system is compatible with 3D FloTrix® vivaSPIN PRO's online live cell counting system, which can precisely monitor the status of viable cells in real time and automatically generate cell proliferation curve, making cell culture process much easier!
Combining CytoNiche's microcarrier and 3D FloTrix® vivaSPIN Automation Bioreactors, the Vero cell culture can achieve a high density of 1×107/mL.
Vero Cells (African green monkey kidney cell)
Vero cells are anchorage-dependent aneuploid cells isolated from the kidney of African green monkeys in 1962. In 1987, the World Health Organization (WHO) officially approved the continuous cell line as the raw material for the production of biological products. Since then, Vero cells have gradually been approved for the production of human vaccines, such as inactivated poliovirus vaccine, oral poliovirus vaccine, and rabies virus vaccine. Since the outbreak of COVID-19 in early 2020, Vero cells have been preferred by many companies around the world for the development and production of the COVID-19 vaccine.
Deng Tao, Nian Xuanxuan, Zhang Jiayou, Huang Shihe, Yang Xiaoming. Development and Application of New Coronary Virus Inactivated Vaccine [J]. Chinese Journal of Biological Laboology, 2021,34 (07): 761-769.
Vero cells have strong adhesive ability and can easily adapt to the microcarrier culture process.
CytoNiche's microcarrier enables continuous amplification and subculture in multistage bioreactors. The peak of cell density can reach 1×107/mL, making it easy to achieve process scale-up.
Comparison of Vero cells' growth on CytoNiche's microcarrier (left) and traditional spherical microcarrier (right). The scale is 250 μm.
By using CytoNiche's microcarrier to culture Vero cells and harvest the viruses: the virus titer of virus A is nearly 2 magnitudes higher than that of traditional 2D technology, while virus B and virus C's virus production rate is also significantly higher than that of traditional microcarrier.
Human diploid cells are primary mesenchymal cells derived from normal human fetal tissue. Compared with animal cells, human diploid cells have no potential carcinogenicity and no animal-derived contaminants. They are ideal raw materials for biological products. However, its long cell doubling time and limited serial passage making it difficult to realize scale-up culture and limit its practical application.
Human diploid cells are widely used in the production of inactivated vaccines such as Hepatitis A, Chickenpox, Poliomyelitis and Rabies virus, as well as Live Attenuated Rubella vaccines; In addition, they are also used in the production of Fibroblasts Interferon and other products.
By using CytoNiche's microcarriers to culture human diploid cell line, the number of cells per gram of microcarrier is 3 - 4 times that of traditional microcarriers during harvest stage, and the cell density can reach 2×106/mL.
Comparison of human diploid cells' growth on CytoNiche's microcarrier (left) and traditional spherical microcarrier (right). The scale is 500 μm.
By using CytoNiche's microcarrier to culture type human diploid cells and harvest the viruses, the antigen content of virus E is about 3 times that of 2D process.
MDCK Cells (Dog-Renal Epithelial Cells)
MDCK cells are anchorage-dependent epithelial cells isolated from the kidney of Cocker Spaniel in 1958. Flucelvax® was approved by FDA in 2012, it was the first vaccine derived from cell culture technology in American, replacing the traditional process based on embryonated chicken eggs which used MDCK cells.
CytoNiche's microcarrier enables continuous amplification and subculture of MDCK cells in multistage bioreactors. The peak of cell density can reach 1×107/mL before seeding viruses.
By using CytoNiche's microcarrier to culture MDCK cells and harvest the viruses, the hemagglutination titer of virus D is 4 times that of traditional spherical microcarrier.
The R&D team of CytoNiche Biotechnology will always be committed to providing an overall solution for the large-scale customized expansion process of cells based on 3D microcarriers to solve your problems encountered during upstream process development of cell and gene therapy, extracellular vesicles, vaccines and protein products, etc.
For more details, please fill in the questionnaire and leave your contact information. We will arrange technicians to contact you!
3D Porous Degradable Microcarriers for Virus Production Solutions Information Application Form
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