Abstract
Veterinary viral vaccines generally comprise either attenuated or chemically inactivated viruses which have been propagated on mammalian cell substrates or specific pathogen free (SPF) eggs. New generation vaccines include chemically inactivated virally-infected whole cell vaccines. The NM57 cell line is a bovine nasal turbinate persistently infected (non-lytic infection) with a strain of the respiratory syncytial virus (RSV). The potential of microcarrier technology for the cultivation in bioreactors of this anchorage dependent cell line for RSV vaccine production has been investigated. Both Cytodex 3 and Cultispher S microcarriers proved most suitable from a selection of microcarriers as growth substrates for this NM57 cell line. Maximum cell densities of 4.12×105 cells ml-1and 5.52×105 cells ml-1 respectively were obtained using Cytodex 3 (3 g l-1) and and Cultispher S (1 g l-1) in 5 l bioreactor cultures. The fact that cell growth was less sensitive to agitation rate when cultured on Cultispher S microcarriers, and that cells were efficiently harvested from this microcarrier by an enzymatic method, suggested Cultispher S is suitable for further evaluation at larger bioreactor scales (>5 l) than that described here.
Keywords: anchorage dependent, animal cell, bioreactor, Cultispher S, Cytodex 3, microcarrier, respiratory syncytial virus, vaccine
Full Text
The Full Text of this article is available as a PDF (191.2 KB).
References
- Aunins JG, Croughan MS, Wang DIC, Goldstein JM. Effects of paddle impeller geometry on power input and mass transfer in small scale animal cell culture vessels. Biotechnol Bioeng. 1986;34:1127–1132. doi: 10.1002/bit.260340902. [DOI] [PubMed] [Google Scholar]
- Barteling SJ, Vreewijk J. Development of foot and mouth disease vaccines. Vaccine. 1991;9:75–88. doi: 10.1016/0264-410X(91)90261-4. [DOI] [PubMed] [Google Scholar]
- Chun B-H, Park S-Y, Moon H-M. Production of live attenuated varicella-zoster virus in human embryonic lung cells using microcarrier. Biotechnol Tech. 1996;10:151–156. doi: 10.1007/BF00158937. [DOI] [Google Scholar]
- Clark JM, Hirtenstein MD. Optimizing culture conditions for the production of animal cells in microcarrier culture. Ann NY Acad Sci. 1981;369:33–46. doi: 10.1111/j.1749-6632.1981.tb14175.x. [DOI] [PubMed] [Google Scholar]
- Croughan MS, Hamel J-FP, Wang DIC. Effects of microcarrier concentration in animal cell culture. Biotechnol. Bioeng. 1988;32:957–982. doi: 10.1002/bit.260320805. [DOI] [PubMed] [Google Scholar]
- Croughan MS, Wang DIC. Growth and death in overagitated microcarrier cell cultures. Biotechnol Bioeng. 1989;33:731–744. doi: 10.1002/bit.260330611. [DOI] [PubMed] [Google Scholar]
- Duvar S, Muthing J, Mohr H, Lehmann J. Scale-up cultivation of primary human umbilical vein endothelial cells on microcarriers from spinner vessels to bioreactor fermentation. Cytotechnology. 1991;21:61–72. doi: 10.1007/BF00364837. [DOI] [PubMed] [Google Scholar]
- Fenner F, Bachmann PA, Gibbs EPJ, Murphy FA, Studdert MJ, White DO. Veterinary Virology. London: Academic Press, Inc.; 1987. [Google Scholar]
- Fleischaker RJ, Sinskey AJ. Oxygen demand and supply in cell culture. Eur J Appl Microbiol Biotechnol. 1981;12:193–197. doi: 10.1007/BF00499486. [DOI] [Google Scholar]
- Hayle AJ. Culture of respiratory syncytial virus infected diploid bovine nasal mucosa cells on Cytodex 3 microcarriers. Arch Virol. 1986;89:81–88. doi: 10.1007/BF01309881. [DOI] [PubMed] [Google Scholar]
- Holland JJ, Villarreal LP, Welsh RM, Oldstone MBA, Kohne D, Lazzarini R, Scolnick E. Long-term persistent vesicular stomatitis virus and rabies infection of cells in vitro. J Gen Virol. 1976;33:193–211. doi: 10.1099/0022-1317-33-2-193. [DOI] [PubMed] [Google Scholar]
- Howard CJ, Stott EJ, Thomas LH, Taylor G, Gourlay RN. Protection against respiratory disease in calves induced by vaccines containing respiratory syncytial virus, parainfluenza type 3 virus, Mycoplasma bovis and M dispar. Vet. Record. 1987;121:372–376. doi: 10.1136/vr.121.16.372. [DOI] [PubMed] [Google Scholar]
- Long B. Animal Cell Biotechnology. London: Academic Press Ltd.; 1994. The Production and Regulation of Veterinary Virus Vaccines; pp. 281–313. [Google Scholar]
- Miller WM, Wilke CR, Blanch HW. Transient responses of hybridoma metabolism to changes in the oxygen supply rate in continuous culture. Bioprocess Eng. 1988;3:101–111. doi: 10.1007/BF00369335. [DOI] [Google Scholar]
- Miller WM, Wilke CR, Blanch HW. Transient responses of hybridoma cells to nutrient additions in continuous culture: I. Glucose pulse and step changes. Biotechnol Bioeng. 1989;33:477–486. doi: 10.1002/bit.260330413. [DOI] [PubMed] [Google Scholar]
- Montagnon BJ, Vincent Falguet JC, Janget B. Thousand litre scale microcarrier culture of Vero cells for killed polio virus vaccine. Promising results. Dev Biol Stand. 1983;55:37–42. [PubMed] [Google Scholar]
- Ng Y-C, Berry JM, Butler M. Optimization of physical parameters for cell attachment and growth on macroporous microcarriers. Biotechnol Bioeng. 1996;50:627–635. doi: 10.1002/(SICI)1097-0290(19960620)50:6<627::AID-BIT3>3.0.CO;2-M. [DOI] [PubMed] [Google Scholar]
- Ohlson S, Branscomb J, Nilsson K. Bead-to-bead transfer of Chinese hamster ovary cells using macroporous microcarriers. Cytotechnology. 1994;14:67–80. doi: 10.1007/BF00772197. [DOI] [PubMed] [Google Scholar]
- Onuma M, Hodatsu T, Yamamoto S, Higashihara M, Masu S, Mikami T, Izawa H. Protection by vaccination against bovine leukemia virus infection in sheep. Am J Vet Res. 1984;45:1212–1215. [PubMed] [Google Scholar]
- Pringle CR, Shirodaria PV, Cash P, Chiswell DJ, Malloy P. Initiation and maintenance of a persistent infection by respiratory syncytial virus. J Virol. 1978;28:199–211. doi: 10.1128/jvi.28.1.199-211.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reitzer LJ, Wice BM, Kennel D. Evidence that glutamine, not sugar, is the major energy source for cultured HeLa cells. J Biol Chem. 1979;254:2669–2676. [PubMed] [Google Scholar]
- Rima BK, Martin SJ. Persistent infection of tissue culture cells by RNA viruses. Med Microbiol Immunol. 1976;162:89–118. doi: 10.1007/BF02121320. [DOI] [PubMed] [Google Scholar]
- Stott EJ, Thomas LH, Taylor G, Collins AP, Jebbett J, Crouch S. A comparison of three vaccines against respiratory syncytial virus in calves. J Hyg. 1984;93:251–261. doi: 10.1017/S0022172400064779. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor G, Stott EJ, Bew M, Fernie BF, Cote PJ, Collins AP, Hughes M, Jebbett J. Monoclonal antibodies protect against respiratory syncytial virus infection in mice. Immunology. 1984;52:137–142. [PMC free article] [PubMed] [Google Scholar]
- Thomas LH, Gourlay RN, Stott EJ, Howard CJ, Bridger JC. A search for new microorganisms in calf pneumonia by the inoculation of gnotobiotic calves. Res Vet Sci. 1982;33:170–182. doi: 10.1016/S0034-5288(18)32331-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van der Velden-de Groot CAM. Microcarrier technology, present status and perspective. Cytotechnology. 1995;18:51–56. doi: 10.1007/BF00744319. [DOI] [PubMed] [Google Scholar]
- Van Wezel AL, Van der Velden-de Groot CAM, Van Herwaarden JAM. The production of inactivated polio vaccine on serially cultivated kidney cells from captive bred monkeys. Dev Biol Stand. 1980;46:151–158. [PubMed] [Google Scholar]
- Varani J, Josephs S, Hillegas WJ. Human diploid fibroblast growth on polystyrene microcarriers in aggregates. Cytotechnology. 1996;22:111–117. doi: 10.1007/BF00353930. [DOI] [PubMed] [Google Scholar]
- Zielke HR, Ozand PT, Tildon JT, Sevdalian DA, Cornblath M. Reciprocal regulation of glucose and glutamine utilisation by cultured diploid fibroblasts. J Cell Physiol. 1978;95:41–48. doi: 10.1002/jcp.1040950106. [DOI] [PubMed] [Google Scholar]