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Master's Dissertation
DOI
10.11606/D.11.2018.tde-20181127-155425
Document
Author
Full name
Dayse Maria de Magalhaes Figueiredo
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1988
Supervisor
Title in Portuguese
Desenvolvimento de metodologia para análise dos componentes de meios de cultura utilizados na produção industrial de vacinas
Keywords in Portuguese
MEIOS DE CULTURA
PRODUÇÃO INDUSTRIAL
VACINAS
Abstract in Portuguese
As vacinas virais, que representam os principais produtos obtidos a partir de células animais, continuam sendo produzidas com partículas virais obtidas a partir de culturas de células em grande escala. A produção industrial da vacina contra febre aftosa envolve o crescimento, em suspensão líquida, de células BHK-21 ("baby hamster kidney cells") usadas como hospedeiras para multiplicação do vírus. Essas células são normalmente cultivadas em um meio nutritivo (Eagle's medium Glasgow modified - MGM) suplementado com soro animal e/ou outros suplementos quimicamente indefinidos como caIdo tripticaseina fosfato (CTF) e hidrolisado de lactoalbumina. A função desses suplementos é estimular o crescimento das células para assegurar a manutenção continua da cultura. O presente trabalho foi realizado com o objetivo de desenvolver uma metodologia prática, pouco dispendiosa e em escala de laboratório, capaz de avaliar com rapidez e confiabilidade, a atividade promotora de crescimento dos meios de cultura utilizados na fabricação industrial de vacinas. A determinação do tempo de dobramento (TD) e da densidade máxima ou desaturação (OS através de curvas de crescimento em monocamada mostrou-se um método bastante eficaz para avaliar os níveis de atividade promotora de crescimento de diferentes meios de cultura, soros e CTF usados no cultivo de células BHK-21. Os resultados obtidos em monocamada se aplicam a células crescendo em suspensão líquida e reproduzem, com excelente aproximação, aqueles obtidos na indústria. Além de permitir o estudo dos fatores promotores de crescimento, esta metodologia oferece perspectivas concretas no sentido da eliminação do soro no cultivo de células BHK-21.
Title in English
Development of a methodologye for the analysis of culture media components used in vaccines' industrial production
Abstract in English
Viral vaccines, the main product obtained from animal cells, have long been produced with viral particles obtained from large scale cell culture. The industrial production of the foot-and- mouth disease (FMD) vaccine involves growth, in liquid suspension of baby hamster kidney (BHK) cells used as the host for viral replication. Normally, these cells are cultured in nutrient medium (Eagle's medium Glasgow modified, MGM) supplemented with animal serum and/or other chemically undefined supplements like tryptose phosphate broth (TPB) and lactalbumin hydrolysate. These supplements stimulate cell growth and thus warrant continuous maintenance of the culture. The objective of the present work was to develop a practical unexpensive method to rapid and reproducibly evaluate the growth promoting activity of culture media components utilized in the industrial production of vaccines, in laboratory scale. Determination of the doubling time (TD) and saturation density (DS) through growth curves is shown to be an efficient and adequate method to evaluate the leveIs of growth promoting activity displayed by the different culture media, sera and TPBs utilized in BHK-21 cells culture. The results obtained in monolayer are shown to directly apply to suspension cultures both in laboratory and in industrial scale. In addition to allowing the study of growth promoting factors, this methodology offers concrete perspectives for the complete elimination of serum as a medium supplement in BHK-21 cultures.
 
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Publishing Date
2018-11-27
 
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