Establishment and Characterization of Three Cell Lines from Aedes triseriatus (Diptera: Culicidae)

Charpentier, G.; Belloncik, S.; Ducros, G.; Fontenille, Didier; Tian, Lingmin; Quiot, J.M.

doi:10.1093/jmedent/32.6.793

Cited by 12 publications

(14 citation statements)

References 10 publications

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“…Regarding the karyological analysis, the number of chromosomes observed matched previous reports on other cell lines of the Culicidae family, including species from the Culex genus (Hsu et al 1970, Charpentier et al 1995, Athawale et al 2002, Sudeep et al 2009). According to Kitzmiller (1976), the number (2n = 6) and morphology of the Culicidae chromosomes are highly conserved among the whole family.…”

Section: Discussionsupporting

confidence: 84%

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Establishment and characterisation of a new cell line derived from Culex quinquefasciatus (Diptera: Culicidae)

Segura

Santamaría

Cabrera

et al. 2012

Mem. Inst. Oswaldo Cruz

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Section: Discussionsupporting

confidence: 84%

“…The occurrence of vesicles in the cell cultures of insects and other arthropods is very common and has been demonstrated in several studies (Charpentier et al 1995, Rey et al 2000, Silva et al 2008). …”

Section: Discussionmentioning

confidence: 85%

Establishment and characterisation of a new cell line derived from Culex quinquefasciatus (Diptera: Culicidae)

Segura

Santamaría

Cabrera

et al. 2012

Mem. Inst. Oswaldo Cruz

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“…En forma similar, en trabajos previos, se ha registrado la heterogénea morfología celular en la iniciación de cultivos in vitro en esta misma especie (8)(9)(10), al igual que en otros mosquitos (14,17,22,23,45). Sin embargo, en los cultivos celulares de insectos, después de muchos subcultivos en serie existe la tendencia al predominio de una determinada forma celular, la cual se mantiene indefinidamente en la línea.…”

Section: Discussionunclassified

Características de nuevos cultivos celulares derivados de tejidos embrionarios de Aedes aegypti (Diptera: Culicidae).

Silva¹,

Escovar²,

Bello³

2005

biomedica

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Introducción. Los cultivos celulares de insectos son una metodología útil en estudios biomédicos y tecnológicos. Objetivo. El propósito principal del presente trabajo fue obtener y caracterizar cultivos celulares derivados de tejidos embrionarios de Aedes aegypti. Materiales y métodos. Se emplearon huevos embrionados para los explantes de tejidos en los medios de cultivos MM/VP12 y L-15/Grace, con suplemento de 20% de suero fetal bovino y una mezcla al 1% de antibiótico y antimicótico, con un rango de pH entre 6,8 y 7,0. Los cultivos se incubaron a una temperatura de 28 o C sin atmósfera de CO 2 . Resultados. El crecimiento celular se obtuvo en el medio L-15/Grace, 3 semanas después de haber sido sembrados los tejidos embrionarios; sin embargo, se necesitaron 6 meses para la formación de la monocapa confluente. Desde agosto de 2003 hasta junio de 2004, se habían realizado 28 subcultivos. Las células se caracterizaron morfológicamente; predominaron las formas epitelioides en subcultivos de pases altos. También se reconocieron las particularidades morfométricas del cariotipo y, además, se determinaron los perfiles isoenzimáticos y moleculares de los cultivos celulares, los cuales se compararon con muestras de adultos de la especie tomadas de la misma colonia y con líneas celulares derivadas de otros insectos. Discusión. Estas células representan, potencialmente, un importante sistema in vitro en investigaciones básicas y aplicadas. Palabras clave: Aedes aegypti, cultivos celulares, vesículas, cariotipos, isoenzimas, RAPD-PCR.Characteristics of new cell cultures derived from embryonic tissues of Aedes aegypti (Diptera: Culicidae) Introduction. Cell cultures from insects are a useful methodology in technological and biomedical studies.Objective. The present work was aimed at obtaining and characterizing cell cultures derived from Aedes aegypti embryonic tissues. Materials and methods. Embryonated eggs were used for embryonic tissue explants in L-15/ Grace and MM/VP12 culture media, supplemented with 20% fetal bovine serum and a mixture of 1% antimycotic and antibiotics, at a pH ranging from 6.8 to 7.0. The incubation temperature was 28 o C; a CO 2 atmosphere was not required. Results. Cell growth was obtained in L-15/Grace medium three weeks after embryonic tissues explants. Six months were required for achieving a confluent monolayer. Twenty-eight serial cell subcultures were carried out from August 2003 to June 2004. Cell morphology was characterized as epithelial in the later subcultures. Karyotype morphometry as well as molecular and isozymatic profiles were established. The cultures were compared with adult samples from the species taken from the same colony and with cell lines derived from other insects.

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“…La presencia de dos grupos diferentes de células en los cultivos celulares derivados del mosquito A. aegypti se debe, posiblemente, a los diversos tipos de tejidos disponibles, a partir de huevos embrionados utilizados durante la formación de los cultivos primarios (Charpentier et al 1995, Rey et al 2000. Sin embargo, predominaron las formas de apariencia epitelioides, como lo registró Ardila et al (2005), diferente a lo observado en las líneas celulares Mos.…”

Section: Discussionunclassified

“…Las vacuolas que fueron apreciadas en las electromicrografías, son similares a las observadas en el citoplasma de una línea celular de A. triseriatus (Charpentier et al 1995). El resultado positivo a la coloración de PAS concuerda con lo descrito en una línea celular de A. eucalypti, en donde se menciona la presencia de vesículas densas, las cuales fueron PAS positivo.…”

Section: Discussionunclassified

Morfología y citoquímica de cultivos celulares de Aedes aegypti (Diptera: Culicidae) y susceptibilidad a Leishmania panamensis (Kinetoplastida: Trypanosomatidae)

Miranda¹,

Sarmiento²,

Caldas³

et al. 2007

RBT

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Morphology and cytochemistry of Aedes aegypti's cell cultures (Diptera: Culicidae) and susceptibility to Leishmania panamensis (Kinetoplastida: Trypanosomatidae). The first cellular line of Aedes aegypti was developed by Grace in 1966; afterwards, other cellular lines of this species have been generated. These have been used for the study of pathogenic organisms like viruses, bacteria and parasites, which demonstrates their importance in biomedical applications. This research describes, for the first time, some cytochemical characteristics of A. aegypti cell cultures, that were infected with (MHOM/CO/87CL412) strain of Leishmania panamensis. A morphological study of the cell culture was also carried out. Maintenance of the cell culture, parasites and infection in vitro were carried out in the Laboratory of Entomology, Cell Biology and Genetics of the Universidad de La Salle. The cell cultures infected with the parasite were maintained in a mixture of mediums Grace/L15, supplemented with 10 % fetal bovine serum (FBS) at pH 6.8 and a temperature of 26 ºC, during 3, 6 and 9 post-infection days. After this, these cell cultures were processed through High Resolution Light Microscopy (HRLM) and Transmission Electron Microscopy (TEM) based on standard protocols defined by the Group of Microscopy and Image Analyses of the Instituto Nacional de Salud. Semi-fine slices of 1 µm colored with toluidine blue were used for the morphological analysis of the culture, and ultra fine cuts of 60 to 90 nm stained with uranyl acetate and lead citrate where used for the ultrastructural study. In addition, PAS and peroxidase staining was carried out in cells fixed with methanol. The morphometric study was analyzed with software ImageJ (NIH). In the semi-fine slices, small cells were observed showing fibroblastic appearance 10.84±2.54 µm in length and 5.31±1.26 µm wide; other cells had epithelial appearance with a great peripheral nucleus, voluminous and vacuolated cytoplasm, 23.04±4,00 µm in length and 13.96±3.70 µm wide. These last ones predominated over the ones with fibroblastic appearance. Regarding the PAS coloration, 7.08 % of the cells presented abundant PAS positive cytoplasmatic granules which indicated polysaccharides presence. The peroxidase test gave a negative result. The greatest percentage of infection (18.90 %) of one total of 101 cells, turned up by day 6. Some cells analyzed by TEM presented a vacuolated aspect cytoplasm; some contained parasites, other fibrillar material and others were empty. The results indicate that A. aegypti cell culture can support the internalization and transformation of the parasite, which demonstrates the capacity that these cell cultures have to be infected with L. panamensis and to maintain the infection for approximately one week. Rev. Biol. Trop. 56 (2): 447-458. Epub 2008 June 30.

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Establishment and Characterization of Three Cell Lines from Aedes triseriatus (Diptera: Culicidae)

Cited by 12 publications

References 10 publications

Establishment and characterisation of a new cell line derived from Culex quinquefasciatus (Diptera: Culicidae)

Establishment and characterisation of a new cell line derived from Culex quinquefasciatus (Diptera: Culicidae)

Características de nuevos cultivos celulares derivados de tejidos embrionarios de Aedes aegypti (Diptera: Culicidae).

Morfología y citoquímica de cultivos celulares de Aedes aegypti (Diptera: Culicidae) y susceptibilidad a Leishmania panamensis (Kinetoplastida: Trypanosomatidae)

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