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Application of 3D Porous Degradable Microcarriers in Vaccine Field - Vero Cells

Application of 3D Porous Degradable Microcarriers in Vaccine Field - Vero Cells

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  • Time of issue:2022-03-31
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(Summary description)Vaccines are produced from bacteria or viruses by artificially using methods such as inactivation, attenuation and genetic recombination to make them non-pathogenic or less pathogenic without affectin

Application of 3D Porous Degradable Microcarriers in Vaccine Field - Vero Cells

(Summary description)Vaccines are produced from bacteria or viruses by artificially using methods such as inactivation, attenuation and genetic recombination to make them non-pathogenic or less pathogenic without affectin

  • Categories:Company News
  • Author:
  • Origin:
  • Time of issue:2022-03-31
  • Views:0
Information

Vaccines are produced from bacteria or viruses by artificially using methods such as inactivation, attenuation and genetic recombination to make them non-pathogenic or less pathogenic without affecting antigenic activity. After vaccination, the human body can develop corresponding immunity to avoid infection.
Vero cells grow adherently and are susceptible to many viruses, and this broad susceptibility has prompted the development of corresponding Vero cell-based vaccines. The bioreactor and microcarrier suspension culture technology enable large-scale cell culture, which has been widely used in vaccine production. For example, inactivated polio vaccine, inactivated COVID-19 vaccine and rabies virus vaccine, etc. To this day, a variety of Vero cell-based vaccines have been marketed. 

Introduction of Vero Cell Culture Process using 3D TableTrix® Microcarriers

The microcarrier suspension culture technology can be carried out in a stirred bioreactor. The 3D TableTrix® microcarrier enables large-scale expansion of cells in the 3D FloTrix® miniSPIN bioreactor (Figure 1). When used in combination with the 3D FloTrix® vivaSPIN automated bioreactor (Figure 2), it can achieve the stepwise scale-up of cells, and complete the harvesting of cells, viruses and cell products. Cell culture process parameters such as temperature, dissolved oxygen, pH, rotating speed, Air, O2, CO2 and N2 can be controlled with high precision and automation, providing a good environment for cells and promoting cell proliferation.

For MDCK cells, continuous scale-up subculture can be achieved in a multi-stage reactor, and the peak cell density can reach nearly 1×107/mL before seeding viruses;

When culturing human diploid cell lines, the cell viability and amplification effect are significantly better than those of traditional microcarriers. When harvesting, the number of cells per gram of microcarriers is 3-4 times that of traditional microcarriers, and the cell density can reach 2×106/mL.

 

 Figure 1: 3D FloTrix® miniSPIN Bioreactor Figure    2: 3D FloTrix® vivaSPIN Automated Bioreactor 

The online live cell counter can monitor the number of viable cells within the system in real time during the culture process (Figure 3). Based on the remote operating system, it can make the experiment more convenient and faster. Combined with mature automatic medium exchange, perfusion and other processes, high-density cell proliferation can be achieved within limited time and space.

Figure 3: Online Cell Density Proliferation Curve 

Derived from the transformation of the scientific research achievements of Tsinghua University, the 3D TableTrix® microcarriers pioneered by CytoNiche have shown great advantages in this field. CytoNiche launched the first 3D Porous Degradable Microcarriers for the viral biological product industry - 3D TableTrix® microcarrier (V series), which can not only be used for the breeding, harvesting and subculturing of adherent cells, but also for virus production; compared with traditional microcarrier, it has obvious advantages in cell expansion and virus production.

Figure 4: 3D TableTrix® Microcarrier (V Series)

Product Features

1. It has a honeycomb-like porous structure, with a pore size range of 30–50 μm and a porosity of >90%;

2. The particle size range is 50–500 μm and can be customized;

3. The specific surface area is more than 9000cm2/g (about 2–3 times the surface area of traditional microcarriers), which can provide more effective growth space and hold more adherent cells under the same feeding density;

4. It has bionic biomechanical characteristics, which can effectively buffer the collision between the fluid shear force and the microcarriers in the stirred reactor, and improve the cell viability, thereby increasing the virus yield;

5. The microcarrier is completely dissolvable, and the lysis carryover testing method for microcarrier (kit) verified by the National Institutes for Food and Drug Control is also provided.

lision between the fluid shear force and the microcarriers in the stirred reactor, and improve the cell vi

 

Process Flow

1. Preliminary Preparation

3D TableTrix® microcarriers (V series) are soaked in PBS in advance and then autoclaved at 121 °C for 30 min before use.

2. Cell Culture

(1) Two-dimensional cells are harvested and seeded into SF500 mL double-arm spinner flasks for breeding in the form of microcarrier suspension culture to obtain a sufficient number of cells.

(2) The cells are digested from the microcarriers and transferred to a 5L-3D FloTrix® vivaSPIN automated bioreactor for culture. The seeding density is 1×106cells/mL, the amount of microcarriers is about 6g/L, and the culture system is 4L. During the culture process, the PECALS™ control system of 3D FloTrix® vivaSPIN is used for automatic medium exchange. The volume of medium replacement is 2L every 24h.

(3) During the culture process, the cell density can be monitored in real time on the PECALS™ system directly through the CELL PRO online cell counter, or the number of cells can be measured by sampling regularly.

3. Virus Production Stage

(1) When a certain cell density is reached, the Vero cell medium is exchanged to virus maintenance solution and the virus is seeded.

(2) Based on the virus production cycle, the supernatant is taken for extracellular virus harvesting or the cells are harvested for intracellular virus harvesting at the proper time.

Figure 5: Schematic Diagram of Vero Cell Expansion

 

【Result Analysis

Using 3D TableTrix® microcarriers and 3D FloTrix® vivaSPIN automated bioreactor, with 4 g/L–10 g/L of microcarriers, a high cell culture density of 1×107cells/mL can be achieved after continuous culture for one week. Vero cells are well attached to microcarriers, and the cell density is sufficient for mass production of vaccines. The relevant research data have shown that their virus production efficiency is significantly higher than that of traditional microcarriers. Other research data are still being updated.

 

【Application of 3D TableTrix® Microcarriers in Serum-Free Culture of Vero Cells】

Through a large number of experimental studies, it was found that after changing to serum-free medium during virus production, the cells still had an obvious tendency to proliferate. This also verifies the advantages of the unique three-dimensional biomimetic structure of 3D TableTrix® microcarriers, which can effectively reduce the need for culture medium of cells and maintain cell proliferation for a period of time.

Conclusion

3D TableTrix® microcarriers (V series) have shown great advantages in the culture of Vero cells, which are superior to the existing microcarrier products on the market. The large-scale expansion of cells can be achieved using microcarriers at relatively low density. And microcarriers can be degraded by digestive enzymes, so that the cells are harvested without damage. In general, using 3D TableTrix® microcarriers (V series), and combined with the unique advantages of 3D FloTrix® vivaSPIN automated bioreactors, the desired number of cells can be harvested in a short time, which greatly meets the needs of scientific research and production, provides new impetus to vaccine companies and has become a better choice.

Process Flow

1. Preliminary Preparation

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Copyright: Beijing CytoNiche Biotechnology Co., Ltd.
Copyright: Beijing CytoNiche Biotechnology Co., Ltd.