The Growth Study of Vero Cells in Different Type of Microcarrier

Yusilawati Ahmad Nor, Nurul Hafizah Sulong, Maizirwan Mel, Hamzah Mohd Salleh, Iis Sopyan

.

DOI: 10.4236/msa.2010.15038                

Abstract

The fact of microcarrier (MC) culture introduces new possibilities and makes possible the practical high-yield culture of anchorage-dependent cells has generated a considerable focus in this study. The objective of this research was to study the comparison of Vero cell growth on different types of commercial microcarriers; Cytodex-1, Cytodex-3, Hil-lex® II and Plastic Plus in spinner vessel and two liters bioreactor cultured for 96 hours. Biological performance of the microcarrier in RPMI media showed the preference of Vero cell grew on Cytodex 3 microcarriers with highest maximum viable cell number (2.4 × 105 cells/ml) followed by Cytodex 1, Hillex and Plustic Plus. Vero cell on Cyto-dex-3 data in spinner flask was compared in bioreactor and result showed higher viable cell number in biorector. Thus, this dextran-crosslink gelatin microcarrier (Cytodex 3) provided the best surface for cell attachment and fast proliferation. At the end of this cell growth improvement will be used for virus transfection producing a vaccine in bioreactor.

Keywords

Cytodex 3, Microcarrier, Spinner Flask, Vero Cells, Vaccines

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	R. Sheet, “History and Characterization of the Vero Cell Line, Technical Report,” Vaccines and Related Biological Products Advisory Committee, 2000.
[2]	K. Trabelsi, S. Rourou, H. Loukil, S. Majoul and H. Kalle, “Comparison of Various Culture Modes for the Production of Rabies Virus by Vero Cells Grown on Microcarriers in a 2L Bioreactor,” Enzyme and Microbial Technology, Vol. 36, No. 4, 2005, pp. 514-519.
[3]	S. Rourou, V. D. Ark, T. B. D. Velden and H. Kallel, “A Microcarrier Cell Culture Process for Propagating Rabies Virus in Vero Cells Grown in a Stirred Bioreactor under Fully Animal Component Free Conditions,” Vaccine, Vol. 25, No. 19, 2007, pp. 3879-3889.
[4]	O. Kistner, M. K. Howard, M. Spruth, W. Wodal, P. Bruhl, M. Gerencer, A. Brian, H. D. Crowe, I. Livey, M. Reiter, I. Mayerhofer, C. Tauer, L. Grillberger, W. Mundt, F. G. Falkner and N. P. Barrett, “Cell Culture (Vero) Derived Whole Virus (H5N1) Vaccine Based on Wild-Type Virus Strain Induces Cross-Protective Immune Respon- ses,” Vaccine, Vol. 25, No. 32, 2007, pp. 6028-6036.
[5]	H. Huang, X. Yi and Y. Zhang, “Improvement of Vero Cell Growth in Glutamate-Based Culture by Supplement- ing Ammoniagenic Compounds,” Process Biochemistry, Vol. 41, No. 12, 2006, pp. 2386-2392.
[6]	F. Horaud, “Absence of Viral Sequences in the WHO Vero Cell Bank: A Collaborative Study,” Development of Biological Standardization, Vol. 76, 1992, pp. 43-46.
[7]	M. Butler, “The Basics Animal Cell Culture and Tech- nology,” Oxford University Press, Oxford, 1996.
[8]	C. A. M. Groot, “Microcarrier Technology, Present Status and Perspective,” Cytotechnology, Vol. 18, No. 1-2, 1995, pp. 51-56.
[9]	H. Clark, K. Dai and K. Masashi, “The Design of Polymer Microcarrier Surfaces for Enhanced Cell Growth,” Biomaterials, Vol. 24, No. 23, 1999, pp. 4253-4264.
[10]	J. Malda and C. G. Frondoza, “Microcarriers in the Engineering of Cartilage and Bone,” Trends in Biotechnology, Vol. 24, No. 7, 2006, pp. 299-304.
[11]	A. J. Sinskey, R. J. Fleischaker and M. A. Tyo, “Production of Cell Derived Products: Virus and Interferon,” Annals of the New York Academy of Sciences, Vol. 369, No. 1, 1981, pp. 47-64.
[12]	L. A. White and E. W. Ades, “Growth of Vero E-6 Cells on Microcarriers in a Cell Bioreactor,” Journal of Clinical Microbiology, Vol. 28, No. 2, 1990, pp. 283-286.
[13]	B. L Seal, T. C. Otero and A. Panitch, “Polymeric Bio- materials for Tissue and Organ Regenaration,” Materials Science and Engineering, Vol. 34, No. 4, 2001, pp. 147- 230.
[14]	R. Freshney, “Culture of animal cells,” Wiley and Sons, Hoboken, 1998.
[15]	M. Saltzman and Kyriakides, “Principle of Tissue Engineering,” Elsevier Inc, Houston, 2007.
[16]	Solo Hill Engineering Inc, “Solohill Microcarrier Beads,” 2009. http://www.solohill.com/files/microcarrier_beads.pdf
[17]	S. C. Wu, C. C. Liu and W. C. Lianc, “Optimization of Microcarrier Cell Culture Process for the Inactivated Enterovirus Type 71 Vaccine Development,” Vaccine, Vol. 22, No. 29-30, 2004, pp. 3858-3864.
[18]	Y. Wang and F. Ouyang, “Recycle of Cyto-dex-3 in Vero Cell Culture,” Bioprocess Engineering, Vol. 21, No. 3, 1999, pp. 207-210.