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. 2010:2010:586363.
doi: 10.1155/2010/586363. Epub 2010 Jun 10.

Production of Newcastle disease virus by Vero cells grown on cytodex 1 microcarriers in a 2-litre stirred tank bioreactor

Mohd Azmir Arifin  1 , Maizirwan Mel, Mohamed Ismail Abdul Karim, Aini Ideris
Affiliations

Production of Newcastle disease virus by Vero cells grown on cytodex 1 microcarriers in a 2-litre stirred tank bioreactor

Mohd Azmir Arifin et al. J Biomed Biotechnol. 2010.

Abstract

The aim of this study is to prepare a model for the production of Newcastle disease virus (NDV) lentogenic F strain using cell culture in bioreactor for live attenuated vaccine preparation. In this study, firstly we investigated the growth of Vero cells in several culture media. The maximum cell number was yielded by culture of Vero cells in Dulbecco's Modified Eagle Medium (DMEM) which was 1.93 x 10(6) cells/ml. Secondly Vero cells were grown in two-litre stirred tank bioreactor by using several commercial microcarriers. We achieved the maximum cell concentration about 7.95 x 10(5) cells/ml when using Cytodex 1. Later we produced Newcastle Disease virus in stirred tank bioreactor based on the design developed using Taguchi L4 method. Results reveal that higher multiplicity of infection (MOI) and size of cell inoculums can yield higher virus titer. Finally, virus samples were purified using high-speed centrifugation based on 3( * *)(3-1) Fractional Factorial Design. Statistical analysis showed that the maximum virus titer can be achieved at virus sample concentration of 58.45% (v/v), centrifugation speed of 13729 rpm, and centrifugation time of 4 hours. As a conclusion, high yield of virus titer could be achieved through optimization of cell culture in bioreactor and separation by high-speed centrifugation.

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Figures

Figure 1
Figure 1
Comparison of Vero cell growth in four different culture media. All media were supplemented with 10% fetal bovine serum (FBS).
Figure 2
Figure 2
Comparison of Vero cell growth on different types of microcarrier in a 2 L stirred tank bioreactor.
Figure 3
Figure 3
Cell attachment of Vero cells on four different microcarriers viewed under inverted light microscope at magnification of 20x: (a) Cytodex 1; (b) Cytodex 3; (c) Hillex; (d) Plastic Plus (PP).
Figure 4
Figure 4
Growth of Vero cells on Cytodex 1 microcarrier and production of NDV: (a) cell inoculums: 1.0 × 105 cells/mL, serum concentration: 0.5%, MOI: 2.0; (b) cell inoculums: 0.5 × 105 cells/mL, serum concentration: 2.0%, MOI: 2.0; (c) cell inoculums: 1.0 × 105 cells/mL, serum concentration: 2.0%, MOI: 0.2; (d) cell inoculums: 0.5 × 105 cells/mL, serum concentration: 0.5%, MOI: 0.2. (↑) indicates cell infection.
Figure 5
Figure 5
Monitoring of Cytodex 1 microcarriers during cell growth and NDV production phases of Vero cells grown in a 2 L bioreactor. Cells were observed with an inverted light microscope without staining: (a) day 0, (b) day 2, (c) day 3, and (d) 2 day post infection.
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References

    1. Adene DF. Country report on the management and health problems of rural poultry stock in Nigeria. In: Proceedings of the Centre for Tropical Agriculture (CTA) Seminar on Small Holder Rural Poultry Production; October 1990; Thessaloniki, Greece. pp. 175–182.
    1. Huang Z, Krishnamurthy S, Panda A, Samal SK. Newcastle disease virus V protein is associated with viral pathogenesis and functions as an alpha interferon antagonist. Journal of Virology. 2003;77(16):8676–8685. - PMC - PubMed
    1. Alexander DJ. Newcastle disease and other avian paramyxoviridae infections. In: Saif YM, Barnes HJ, Glisson JR, Fadly AM, McDougald LR, Swayne DE, editors. Diseases of Poultry. 11th edition. Iowa State University Press; 2003. pp. 63–87.
    1. De Leeuw O, Peeters B. Complete nucleotide sequence of Newcastle disease virus: evidence for the existence of a new genus within the subfamily Paramyxovirinae. Journal of General Virology. 1999;80(1):131–136. - PubMed
    1. Seal BS, King DJ, Sellers HS. The avian response to Newcastle disease virus. Developmental and Comparative Immunology. 2000;24(2-3):257–268. - PubMed

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