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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2015.tde-08062015-100249
Document
Author
Full name
Gisele Cristiane de Melo Dias
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Borella, Maria Ines (President)
Batlouni, Sérgio Ricardo
Oliveira, Sergio Ferreira de
Romagosa, Elizabeth
Siviero, Fábio
Title in Portuguese
Caracterização, isolamento e cultura de espermatogônias primárias de curimbatá, Prochilodus lineatus (Valencienes, 1847).
Keywords in Portuguese
Cultura de células
Espermatogônias primárias
Estereologia
Morfometria
Peixes teleósteos
Testículos
Abstract in Portuguese
Machos adultos de P. lineatus tiveram suas gônadas processadas de acordo com as rotinas de microscopia de luz e microscopia eletrônica de transmissão. Para a cultura de células, os testículos foram digeridos enzimaticamente, a suspensão testicular foi separada por gradiente descontínuo com Percoll seguido pelo plaqueamento diferencial por adesão e as células foram cultivadas. Foi realizado o método de enriquecimento das espermatogônias por citometria de fluxo. Os testículos apresentam as regiões anterior, média e posterior com distribuição semelhante dos tipos de células, e o diâmetro nuclear das células germinativas diminui significativamente durante a espermatogênese. As espermatogônias cultivadas por 15 dias com meio para proliferação celular resultaram em grandes aglomerados celulares que foram caracterizados com o anti-Vasa, anti-GFRa1 e anti-OCT4. As culturas que receberam o meio para diferenciação celular mostraram processo de proliferação lento das espermatogônias primárias comparado com a cultura que teve o meio indicado para proliferação celular.
Title in English
Characterization, isolation and culture of primary spermatogonias of curimbatá, Prochilodus lineatus (Valencienes, 1847).
Keywords in English
Cell culture
Morphometry
Primary spermatogonia
Stereology
Teleost fish
Testes
Abstract in English
Adult males of P. lineatus had their gonads used according routines of light microscopy and transmission electron microscopy. For cell culture, the testes were enzymatically digested; testicular suspension was separated by discontinuous gradient with Percoll followed by adhesion differential plating and the cells were cultured. The enrichment of the spermatogonia was carried by flow cytometry. The testes present three regions with similar distribution of cell types, and nuclear diameter of germ cells decreases significantly during spermatogenesis. The spermatogonia cultured for 15 days with medium for cell proliferation, resulted in large cell agglomerates which were characterized with the antibodies anti-Vasa, anti-GFRa1 and anti-anti-OCT4. The cultures that receiving medium for cell differentiation showed slow proliferation process of primary spermatogonia compared to cell culture medium suggestive for cell differentiation.
 
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Publishing Date
2015-06-08
 
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