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Doctoral Thesis
DOI
10.11606/T.87.2016.tde-04122015-152642
Document
Author
Full name
Angelina Cirelli Moraes
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Battesti, Darci Moraes Barros (President)
Morena, Diva Denelle Spadacci
Nakano, Eliana
Onofrio, Valéria Castilho
Piazza, Roxane Maria Fontes
Title in Portuguese
Estabelecimento e caracterização de células embrionárias de Amblyomma sculptum Berlese (Acari: Ixodidae).
Keywords in Portuguese
Amblyomma sculptum
Biomarcadores
Células embrionárias
Cultura de células
Linhagem celular
Patógenos
Abstract in Portuguese
Cultura de células de carrapatos, é uma ferramenta importante, para isolar e estudar os agentes de doenças transmissíveis. Amblyomma sculptum é o principal vetor de Rickettsia rickettsii, o agente da febre maculosa. O objetivo deste estudo foi estabelecer e caracterizar as células deste carrapato, além de testar seu potencial uso, como substrato para o crescimento e isolamento de patógenos. A partir de massas de ovos de A. sculptum, culturas primárias foram, preparadas em meio L-15B, subcultivadas ao atingirem a confluência necessária e criopreservadas em diversas passagens. A boa recuperação celular estabeleceu a linhagem IBU/ASE-16, a qual foi testada para, Rickettsia spp, Trypanosoma theileri e Leishmania infantum chagasi, obtendo-se bons resultados. A citometria de fluxo analisou a expressão de marcadores de células-tronco, proliferação, diferenciação e regulação do ciclo celular, nas IBU/ASE-16. O Δψm mostrou atividade das mitocôndrias e ausência de células inativas. A microscopia confocal, localizou estruturas celulares marcadas com fluorocromos, enquanto que a MEV,mostrou, junções intercelulares, matriz extracelular e redes neuronais. O teste de tumorigênese em camundongos Balb/c nu/nu, não resultou em crescimento de tumores.
Title in English
Establishment and characterization of embryonic cells of Amblyomma sculptum Berlese (Acari: Ixodidae).
Keywords in English
Amblyomma sculptum
Biomarkers
Cell culture
Cell line
Embryonic cells
Pathogens
Abstract in English
Cell cultures of ticks are important tools to isolate and study agents of transmissible diseases. Amblyomma sculptum is the main vector of Rickettsia rickettsii, the agent for spotted fever. The aim of this study was to establish and characterize the cells of this tick, and test their potential use as a substrate for the growth and isolation of pathogens. From a egg masses of A. sculptum, primary cultures were prepared in L-15B medium, subcultured as they reached the necessary convergence and cryopreserved in several passages. The good cell recovery established IBU/ASE-16 lineage, which was tested for Rickettsia spp, Trypanosoma theileri and Leishmania infantum chagasi, yielding good results. Flow cytometry analyzed the expression of stem cell markers, proliferation, differentiation and regulation of the cell cycle, in the IBU/ASE-16 lineage. The Δψm showed activity of mitochondria and absence of inactive cells. Confocal microscopy located cell structures marked with fluorochromes, while MEV showed intercellular junctions, extracellular matrix and neural networks. Tumorigenesis tests in Balb/c nu/nu mice resulted in no tumor growth.
 
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Release Date
2018-03-14
Publishing Date
2016-03-14
 
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