Small ruminant lentiviruses isolated from peripheral blood leukocytes and target organs can be propagated in vitro in fibroblasts derived from goat synovial membrane cells. These cells are obtained from tissues collected from embryos or fetuses and are necessary for the establishment of the fibroblast primary culture. A new alternative type of host cells, derived from goat umbilical cord, was isolated and characterized phenotypically with its main purpose being to obtain cell monolayers that could be used for the diagnosis and isolation of small ruminant lentiviruses in cell culture. To accomplish this goal, cells were isolated from umbilical cords; characterized phenotypically by flow cytometry analysis; differentiate into osteogenic, chondrogenic and adipogenic lineage; and submitted to viral challenge. The proliferation of goat umbilical cord cells was fast and cell monolayers formed after 15 days. These cells exhibited morphology, immunophenotype, growth characteristics, and lineage differentiation potential similar to mesenchymal stem cells of other origins. The goat umbilical cord derived cells stained positive for vimentin and CD90, but negative for cytokeratin, CD34 and CD105 markers. Syncytia and cell lysis were observed in cell monolayers infected by CAEV-Cork and MVV-K1514, showing that the cells are permissive to small ruminant lentivirus infection in vitro. These data demonstrate the proliferative competence of cells derived from goat umbilical cords and provide a sound basis for future research to standardize this cell lineage.
RESUMO -O crescimento e desenvolvimento do rebanho caprino no Nordeste são observados com o aumento na produção da pecuária do Brasil. Esse aumento é reflexo, a princípio, das maiores exigências do mercado consumidor por produtos de melhor qualidade obtidos a partir de rebanhos de alto padrão zootécnico. As pesquisas atuais ilustram a necessidade de dispor de biomarcadores que auxiliem a indicação do potencial reprodutivo dos animais, uma vez que isso não pode ser expresso apenas com o exame andrológico. A avaliação da expressão das proteínas, tomando-as como biomarcadores, é análise potencial uma vez que estas, dentre os constituintes do plasma seminal, são encontradas em maior quantidade na forma de complexos associados, desempenhando papel crucial em todos os processos relacionados à capacidade fecundante dos espermatozoides. Essa análise é realizada por métodos de separação e detecção simultânea de proteínas utilizando técnicas como a eletroforese bidimensional (2DE) ou cromatografias, acoplados a métodos cada vez mais eficientes e sensíveis de identificação e quantificação de níveis de expressão de proteínas por espectrometria de massas. O objetivo desta revisão é abordar sobre a técnica de eletroforese bidimensional associada à espectrometria de massa como ferramenta na análise da expressão de proteínas dentro do campo da proteômica.Palavras-Chave: proteínas, plasma seminal, eletroforese 2DE, espectrometria de massa.ABSTRACT -The growth and development of goat herds in the northeast are being observed due to the increase in livestock production in Brazil. This increase reflects demands of the consumer market for high quality product, which are obtained from flocks of high standard zootechnics. Current research illustrates the need for biomarkers that indicate an animal´s reproductive potential, since this cannot be expressed solely with andrologic evaluation. For this reason, the expression of the proteins as biomarkers is a potential analysis. The proteins from the seminal plasma constituents are found in larger amounts in the form of associated complexes, playing a crucial role in all processes related to the fertilizing capacity of sperm. This analysis is performed by separation methods and simultaneous detection of proteins using techniques such as two-dimensional electrophoresis (2DE) or chromatography. These techniques are coupled with methods to identify and quantify expression levels of proteins by mass spectrometry that are increasingly efficient and sensitive. The aim of this review was to discuss the technique of two-dimensional electrophoresis combined with mass spectrometry as a tool in the analysis of protein expression within the field of proteomics.
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