Egg and fourth instar larvae gut of Aedes aegypti as a source of stem cells

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art_MARIO_Egg_and_fourth_instar_larvae_gut_of_Aedes_2016.PDF (2.23 MB)
Citações na Scopus
2
Tipo de produção
article
Data de publicação
2016
Editora
CHURCHILL LIVINGSTONE
DOI
Indexadores
Scopus
Web of Science
PubMed
Título da Revista
ISSN da Revista
Título do Volume
Autores
MARIO, Lara C.
BORGHESI, Jessica
CRIVELLARI-DAMASCENO, Wilson T.
FAVARON, Phelipe O.
WILL, Sonia E. A. L.
MARIA, Durvanei A.
MIGLINO, Maria A.
Autor de Grupo de pesquisa
Editores
Coordenadores
Organizadores
Citação
TISSUE & CELL, v.48, n.5, p.558-565, 2016
Projetos de Pesquisa
Unidades Organizacionais
Fascículo
Resumo
According to the World Health Organization, 2015 registered more than 1.206.172 cases of Dengue in the Americas. Recently, the Aedes aegypti has been not only related to Dengue, but also with cases of Zika virus and Chikungunya. Due to its epidemiological importance, this study characterized the morphology of the embryonated eggs of A. aegypti and provided a protocol to culture stem cells from eggs and digestive tract of fourth instar larvae in order to examine cell biology and expression of markers in these vectors. Cells were isolated and cultured in DMEM-High at 28 degrees C, and their morphology, cell cycle and immunophenotyping were examined. Morphologically, embryos were at the end of the embryonic period and showed: head, thorax, and abdomen with eight abdominal segments. The embryonic tissues expressed markers related to cell proliferation (PCNA), pluripotency (Sox2 and OCT3/4), neural cells (Nestin), mesenchymal cells (Vimentin and Stro-1), and endosomal cells (GM130 and RAB5). In culture, cells from both tissues (eggs and larvae gut) were composed by a heterogeneous population. The cells had a globoid shape and small size. Cell cycle analysis on passage 1 (P1) showed 27.5% +/- 2.0% of cell debris, 68% of cells on GO-G1 phase, 30.2% on S phase, 1.9% +/- 0.5% on G2-M phase. In addition, cells on passage 2 showed: 10% of cell debris, 92.4% of cells on G0-G1 phase, 6.8% on S phase, 0.6% on G2-M phase. Embryonated eggs expressed markers involved with pluripotency (Sox2 and Oct 3/4), mesenchymal cells (vimentin and Stro-1), neural cells (Nestin), and cellular death by apoptosis (Caspase 3). Specific endosomal markers for insect cells (GM130 and RAB5) were also highly expressed. In cell culture of A. aegypti larvae gut the same labeling pattern was observed, with a small decrease in the expression of mesenchymal (vimentin and Stro-1) and neural (Nestin) markers. In summary, we were able to establish a protocol to culture embryonated eggs and larvae gut of A. aegypti, describing the characteristics of undifferentiated cells, as well as the cell cycle and expression of markers, which can be used for biotechnology studies for the biological control of this vector.
Palavras-chave
Dengue, Cell culture, Morphology, Emerging diseases
URI
https://observatorio.fm.usp.br/handle/OPI/17190
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Coleções
Artigos e Materiais de Revistas Científicas - FM/Outros
Artigos e Materiais de Revistas Científicas - ODS/02
Artigos e Materiais de Revistas Científicas - ODS/03