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Master's Dissertation
DOI
10.11606/D.87.2001.tde-28072005-115307
Document
Author
Full name
Adriana Yurie Yokomizo
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2001
Supervisor
Committee
Pereira, Carlos Augusto (President)
Pradella, Jose Geraldo da Cruz
Suazo, Claudio Alberto Torres
Title in Portuguese
Obtenção de antígeno viral a partir de culturas de células vero em microcarregadores porosos
Keywords in Portuguese
células Vero
Cultispher
cytopore
microcarregadores porosos
vírus rábico
Abstract in Portuguese
Cultivos de células VERO em microcarregadores porosos de celulose (Cytopore) e porosos de gelatina (Cultispher G) com rendimento celular três vezes maior, foram obtidos em relação às culturas em microcarregadores sólidos de DEAE-dextran (Cytodex tipo 1). No Cytopore e Cultispher G atingimos 1.265 e 1.103 células/microcarregador, respectivamente e no Cytodex 1, 256 células/microcarregador. A concentração celular nos suportes porosos Cytopore aumentou 43 vezes o número de células iniciais, 37,5 vezes no Cultispher G e 17 vezes no Cytodex 1. As culturas de células VERO nos microcarregadores porosos foram utilizadas com êxito para a replicação do vírus rábico (PV/VERO), quando infectadas com MOI de 0,05. Títulos de 104,8 FFD50/mL determinados pelo método de inibição de focos fluorescentes (RFFIT) foram obtidos 96 horas após a infecção. Estudos de microscopia óptica e eletrônica de varredura e transmissão para análise da estrutura destes suportes, mostraram boa colonização celular e a eficiência da replicação viral. A padronização de metodologia para cultura e infecção viral de células VERO nos microcarregadores porosos indica a utilidade destes suportes para produção de antígenos e vacinas virais e a potencialidade do sistema para a obtenção de produtos biotecnológicos em geral.
Title in English
Obtention of viral antigens from the Vero cell cultures on porous microcarriers.
Keywords in English
Cultispher
Cytopore
macroporous
rabies virus
Vero cells
Abstract in English
Cultures of VERO cell on porous cellulose-coated microcarriers (Cytopore) and on porous gelatin-coated (Cultispher G) with a cellular yield 3 times higher, was obtained in relation to DEAE-dextran solid microcarriers (Cytodex type 1). In the cultures on Cytopore and Cultispher G we obtained 1.265 and 1.103 cells/microcarrier, respectively and 256 cells/microcarrier on Cytodex 1. The cell concentration for the Cytopore porous suports increased 43 times from the initial cell number, 37,5 times for the Cultispher G and 17 times for the Cytodex 1. The VERO cell cultures on porous microcarriers were efficiently used for the replication of the rabies virus (PV/VERO), when infected with 0,05 MOI. Titers of 104,8 FFD50/mL determined by the fluorescent focus inhibition method (RFFIT) were obtained 96 hours post-infection. Studies on light microscopy and scanning and transmission eletronic microscopy carried out to analyse the suports structure, showed a good cell population and efficiency of virus infection. The standardization of a methodology for culture and virus infection of VERO cells on porous microcarriers indicate the usefulness of these porous supports to the antigens and viral vaccines production and the potenciality of this system for the attainment of biotechnological products.
 
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2005_003.pdf (2.15 Mbytes)
Publishing Date
2007-03-01
 
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