Applications Vaccixcell

Cell Lines

Esco VacciXcell has experience in the culture of different cell lines used for various applications including vaccine development and production, expression of recombinant proteins, and pharmacokinetic studies.

BHK or Baby Hamster Kidney cells are anchorage-dependent cells, originally derived from a 1-day old Syrian golden hamster. BHK-21 cells have been shown to be an efficient system for the production of rabies vaccine for human use, for which Esco VacciXcell’s tide motion principle is currently being used as a production system. Aside from this, Esco VacciXcell’s tide motion system has been also been shown to successfully produce virus-like particles expressed in BHK-21 cell lines through transfection by recombinant baculoviruses.

 

Case Sources:

Existing Customers

Chen, Y., Wu, J., Wang, K., Chiang, Y., Lai, C., Chung, Y., & Hu, Y. (n.d.). Baculovirus-mediated production of HDV-like particles in BHK cells using a novel oscillating bioreactor. Journal of Biotechnology, 135-147.

CHO cells are anchorage-dependent cells, originally derived from the ovary of a female Cricetulus griseus or Chinese hamster. The CHO cell line has been widely used for the expression of different recombinant proteins. Esco VacciXcell’s tide motion bioreactor system has exhibited success in the expression of recombinant proteins, human growth hormones, and monoclonal antibodies from recombinant CHO cells.

 

Case Sources:

Asaoka, Y., Tanaka, T., Tsumoto, K., Tomita, M., & Ide, T. (n.d.). Efficient expression of recombinant soluble human FcγRI in mammalian cells and its characterization. Protein Expression and Purification, 155-161.

Jepson, S., Vought, B., Gross, C., Gan, L., Austen, D., Frantz, J., . . . Krauss, R. (2012). LINGO-1, a Transmembrane Signaling Protein, Inhibits Oligodendrocyte Differentiation and Myelination through Intercellular Self-interactions. Journal of Biological Chemistry, 22184-22195.

Akiyama, M., Nakayama, D., Takeda, S., Kokame, K., Takagi, J., & Miyata, T. (n.d.). Crystal structure and enzymatic activity of an ADAMTS-13 mutant with the East Asian-specific P475S polymorphism. J Thromb Haemost Journal of Thrombosis and Haemostasis, 1399-1406.

MDCK or Madin-Darby Canine Epithelial cells are anchorage-dependent cells that originated from the kidney cells of an adult female Canis familiaris. The MDCK cell line has been used for various applications, including vaccine production and in vitro drug permeability assays. In particular, Esco VacciXcell’s tide motion bioreactor system has been and is currently being used in the development of viral vaccines against H5N1, H7N9, H1N1, H3N2, H3N4, H3N5 and H3N7 for both human and animal uses.  

Esco VacciXcell offers a complete platform for the use of MDCK cell line with tide motion bioreactor system. Since MDCK cell line grows very fast, controlling the optimum pH would be difficult. MDCK cell line performs best using 500AP bottles when doing trial with CelCradle. It minimizes pH control problems and solves media exchange scheme

 

Case Sources:

Existing Customers

Haredy, A., Takenaka, N., Yamada, H., Sakoda, Y., Okamatsu, M., Yamamoto, N., . . . Okamoto, S. (2013). An MDCK Cell Culture-Derived Formalin-Inactivated Influenza Virus Whole-Virion Vaccine from an Influenza Virus Library Confers Cross-Protective Immunity by Intranasal Administration in Mice. Clinical and Vaccine Immunology, 998-1007.

Haredy, A., Yamada, H., Sakoda, Y., Okamatsu, M., Yamamoto, N., Omasa, T., . . . Yamanishi, K. (2014). Neuraminidase gene homology contributes to the protective activity of influenza vaccines prepared from the influenza virus library. Journal of General Virology, 2365-2371.

Vero cells are anchorage-dependent cells, originally derived from the kidney of an adult Cercopithecus aethiops or African green monkey. The Vero cell line has been used for the development of vaccine against various diseases, including yellow fever, influenza, and rabies. In particular, Esco VacciXcell’s tide motion bioreactor system has been and is currently being used in the development of viral vaccines for both human and animal use against Enterovirus 71 (EV71), Infectious Bursal Disease Virus (IBDV), rabies, and Japanese Encephalitis Virus (JEV).

Esco VacciXcell offers a complete platform for the use of the Vero cell line with its tide motion bioreactor system and Plus™ Vero culture medium, ensuring the optimal growth of Vero cells for different applications.

 

Case Sources:

Existing Customers

Toriniwa, H., & Komiya, T. (n.d.). Japanese encephalitis virus production in Vero cells with serum-free medium using a novel oscillating bioreactor. Biologicals, 221-226.

Sf-9 cells originate from the ovary of a female pupa Spodoptera frugiperda or fall armyworm. Sf-9 cells can be grown either in adherent or suspension culture. Esco VacciXcell’s tide motion bioreactor has been used to successfully culture and proliferate Sf-9 cells and as well as to produce recombinant proteins via transfection with baculoviruses.

 

Case Sources:

Existing Customers

Hu, Y., Lu, J., & Chung, Y. (n.d.). High-density cultivation of insect cells and production of recombinant baculovirus using a novel oscillating bioreactor. Cytotechnology, 145-153.

Lu, J., Chung, Y., Chan, Z., & Hu, Y. (n.d.). A Novel Oscillating Bioreactor BelloCell: Implications for Insect Cell Culture and Recombinant Protein Production. Biotechnology Letters Biotechnol Lett, 1059-1065.

HEK-293 cells are anchorage-dependent cells that originate from the embryonic kidney of a human fetus. The HEK-293 cell line has been used for different applications, including drug and toxicity assay, vaccine development, and drug discovery. Particularly, Esco VacciXcell’s tide motion bioreactor system has been used to successfully produce recombinant proteins from recombinant HEK-293 cells and adenovirus production.

HEK-293T must be cultured with BioNOC II coated with PEI (available from Esco VacciXcell).

The following table shows cGMP compliant customers using TideXcell in their projects. Company identities and some other information are hidden due to Non-disclosure agreement (NDA) between the company and VacciXcell.

 

Cell Line Density Product
HEK-293 3.5 x107 108pfu/ml of Adenovirus
HEK-293 3.4x10^9 247 IVP/cell of rAAV
HEK-293 1.4x10^9 3.7 pmoles of radioligandY mg−1 protein

 

Case Sources:

Existing Customers

Huang, K., Lo, W., Chung, Y., Lai, Y., Chen, C., Chou, S., & Hu, Y. (n.d.). Combination of Baculovirus-Mediated Gene Delivery and Packed-Bed Reactor for Scalable Production of Adeno-Associated Virus. Human Gene Therapy, 1161-1170.

Ho, L., Greene, C., Schmidt, A., & Huang, L. (n.d.). Cultivation of HEK 293 cell line and production of a member of the superfamily of G-protein coupled receptors for drug discovery applications using a highly efficient novel bioreactor. Cytotechnology, 117-123.

Tabata, S., Nampo, M., Mihara, E., Tamura-Kawakami, K., Fujii, I., & Takagi, J. (n.d.). A rapid screening method for cell lines producing singly-tagged recombinant proteins using the “TARGET tag” system. Journal of Proteomics, 1777-1785.

Yasui, N., Mihara, E., Nampo, M., Tamura-Kawakami, K., Unno, H., Matsumoto, K., & Takagi, J. (n.d.). Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation. Journal of Immunological Methods, 153-160.

Kato, K., Nishimasu, H., Mihara, E., Ishitani, R., Takagi, J., Aoki, J., & Nureki, O. (2012). Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Enpp1. Acta Cryst Sect F Acta Crystallogr F Struct Biol Cryst Commun Acta Cryst Sect F Struct Biol Cryst Commun Acta Cryst F Struct Biol Cryst Commun Acta Cryst F Acta Crystallographica Section F Structural Biology and Crystallization Communications Acta Crystal, 778-782.

PK-15 cells are anchorage-dependent cells that originated from the kidney of an adult pig (Sus scrofa). The PK-15 cell line has been widely used in vaccine and drug development studies for different pig viruses and diseases. Esco VacciXcell’s tide motion bioreactor system has been used for the successful development and production of vaccine against the hog cholera virus.

 

Case Sources:

Existing Customer

MDBK cells are anchorage-dependent cells that originated from the kidney of an adult male Bos Taurus. The MDBK cell line has been used in vaccine and drug development studies against different diseases including influenza and herpes simplex virus. Esco VacciXcell’s tide motion bioreactor system has been used for the succesful development and production of vaccine against Bovine Herpesvirus (BHV) and other viruses.

 

Case Sources:

Existing Customer

Mesenchymal stem cell (MSC) is a type of pluripotent cell that has the capability of differentiating into various types of cells. MSCs are used for both cell therapy procedures in both autologous and allogeneic applications.

(Sourced from http://www.bioinformant.com/benefits-of-mesenchymal-stem-cells/)

Mesenchymal stem cells (MSCs) are advantageous over other stem cells types for a variety of reasons. First, they avoid the ethical issues that surround embryonic stem cell research.

Second, repeated studies have found MSCs to be immuno-privileged, which make them an advantageous cell type for allogeneic transplantation. MSCs reduce both the risks of rejection and complications of transplantation.

Third, there have been advances in the use of autologous mesenchymal stem cells to regenerate human tissues, including cartilage, meniscus, tendons, and bone fractures, because MSCs can exert regenerative effects through honing to sites of damage, paracrine signalling, regulating the immune response, and affecting the microenvironment.

In combination, these traits make MSCs of intense therapeutic interest, because they represent a population of cells with the potential to treat a wide range of acute and degenerative diseases.

Benefits of MSCs Relative to Other Stem Cells

Interestingly, MSCs have a range of benefits compared to other stem cell types, as presented below:

  1. Well-Characterized: MSCs are a well-characterized population of adult stem cells, with over 36,000 scientific articles published about them.
  2. Non-Controversial: MSCs avoid the ethical issues of embryonic stem cells, as they can be derived from sources that include adult bone marrow and adipose tissue.
  3. Diverse Differentiation Potential: MSCs can form a variety of cell types in the laboratory, including those of both intra- and extra-mesenchymal lineage. These cell types include: fat (adipocytes), bone (osteoblasts), skin (dermal cells), nerve (neural cells), cartilage (chondrocytes), muscle (skeletal myocytes), tendons (tenocytes), marrow stroma, ligaments, and more.
  4. Ease of Growth in Culture: Advanced knowledge exists for how to grow MSCs in culture, including protocols for isolation, expansion, and differentiation.
  5. Flexible Propagation: MSCs can be grown and propagated in culture for extended periods, without losing differentiation potential.
  6. Clinically Relevant Volumes: Unlike many other types of adult stem cells, MSCs can be acquired in the quantities required for clinical applications, as knowledge exists for how to culture the cell type in 3D bioreactors. It is understood that reduced oxygen conditions, along with available nutrients, assist MSC expansion under bioreactor condition.
  7. Role as Regulatory Cells: MSCs synthesize and secrete a variety of macromolecules that are known regulators of hematopoietic and bone-resorbing cells.
  8. Delivery of Gene Products: MSCs can take up exogenous DNA and keep introduced genes, an attribute that may allow use of the cells in therapeutic delivery of molecules to target regions of the body.
  9. Favorable Immune Status: MSCs lack the co-stimulatory molecules of the B7 family that are required to initiate an immune response. This allows the administration of MSC preparations across MHC barriers without concern for immunological rejection or the need for immunosuppression, making MSCs a universal stem cells source.
  10. Commercially Available Research Tools: Currently, dozens of research supply companies offer MSC-based products, making research tools for this cell type easily accessible.

(End of quoted article)

Esco VacciXcell's Tidemotion bioreactor have been proven to be an effective tool for culturing MSC for clinical application. CelCradle™ is a lab scale desktop bioreactor capable of high density adherent cell culture. The CelCradle™ is capable of culturing MSC for four (4) individual patients at the same time without the risk of cross contamination. CelCradle™ is also easily scalable to TideCell®, the pilot/production scale tidemotion bioreactor model. Tidemotion bioreactors are truly linearly scalable up to 5,000L and is the only bioreactor currently available in the market that is able to culture 1 Trillion cells for allogeneic applications.

Sources:

Benefits of Mesenchymal Stem Cells (MSC) Over Other Stem Cell Types www.bioinformant.com

Existing Customer