Culture of mosquito cells in eagle’s medium

Shih, Karen M.; Gerenday, Anna; Fallon, Ann M.

doi:10.1007/s11626-996-0010-1

Cited by 43 publications

(35 citation statements)

References 9 publications

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“…1). An Anopheles cell line, ASEIV, previously suspected of being of gut origin (Kurtti and Munderloh, 1989;Shih et al, 1998;Fallon and Sun, 2001), did not display the same banding profile as that of Anopheles gambiae and An. stephensi midgut lysates (data not shown).…”

Section: Mg96 Identifies a Conserved Oligosaccharide In The Blood-fedmentioning

confidence: 88%

Sugar epitopes as potential universal disease transmission blocking targets

Dinglasan

Valenzuela

Azad

2005

Insect Biochemistry and Molecular Biology

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Section: Mg96 Identifies a Conserved Oligosaccharide In The Blood-fedmentioning

confidence: 88%

Sugar epitopes as potential universal disease transmission blocking targets

Dinglasan

Valenzuela

Azad

2005

Insect Biochemistry and Molecular Biology

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“…The cells were adapted to and subsequently maintained in Eagle's medium, supplemented with non-essential amino acids, glutamine, vitamins, penicillin, and streptomycin, and 20 % heat-inactivated fetal bovine serum, essentially as described previously (Shih et al 1998). This formulation of the medium with 20% serum is called E-20.…”

Section: Cells and Culture Conditionsmentioning

confidence: 99%

“…Because these media contain undefined components such as yeastolate and lactalbumin hydrolysate, which can compromise isotope incorporation and use of metabolic inhibitors, we adapted Aa23 cells to a supplemented version of Eagle's (E) medium containing 20% heat-inactivated, fetal bovine serum (E-20). Apart from the serum, Eagle's medium with additional vitamins, non-essential amino acids, and glutamine (Shih et al 1998) is chemically defined.…”

Section: Growth Characteristics Of Aa23 Cellsmentioning

confidence: 99%

Cytological properties of an Aedes albopictus mosquito cell line infected with Wolbachia strain wAlbB

Fallon

2008

In Vitro Cell.Dev.Biol.-Animal

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In vitro production of the obligate intracellular bacterium, Wolbachia pipientis, is essential to its manipulation as a genetic tool to spread transgenes within vector populations. We have adapted the Wolbachia-infected Aa23 Aedes albopictus mosquito cell line to Eagle's minimal medium, supplemented with nonessential amino acids, glutamine, and 20% fetal bovine serum. When plated at low densities, Aa23E cells grew as patchy monolayers, comprised of non-contiguous clusters of cells that gave rise to solid clumps of tightly adherent cells. Multicellular clumps eventually detached from the substrate and floated freely in the medium. Removal of Wolbachia by treatment with tetracycline did not alter the cytological properties of the host cells, which had a population doubling time of 4-5 d. The presence of Wolbachia was monitored by Giemsa staining of cytological preparations, polymerase chain reaction (PCR) amplification of Wolbachia 16S ribosomal DNA, and by simultaneous PCR amplification of ribosomal protein genes from Wolbachia and mosquito host cell genomes. Wolbachia morphology was pleomorphic, and Wolbachia DNA persisted in the culture medium for several weeks after degradation of PCR-amplifiable host cell DNA.

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“…The Aedes albopictus cell line C7-10 was cultured in Eagle's medium containing 5% fetal bovine serum as described previously (Shih et al, 1998). Cells were maintained at 30°C in a 5% CO 2 atmosphere.…”

Section: Cells and Culture Conditionsmentioning

confidence: 99%

The insect repellent DEET (N,N-diethyl-3-methylbenzamide) increases the synthesis of glutathione S-transferase in cultured mosquito cells

2010

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DEET (N,N-diethyl-3-methylbenzamide) is the active ingredient used in many commonly used insect repellents, but its mode of action remains poorly understood. Efforts to identify properties that could lead to the development of more effective active ingredients have distinguished among DEET's repellent, deterrent, and insecticidal activities. We used an Aedes albopictus mosquito cell line to evaluate DEET's toxicological properties in the absence of sensory input mediated by the olfactory system. When cells were treated with DEET and labeled with [(35)S]methionine/cysteine, a single 25-kDa protein was induced, relative to other proteins, on SDS-polyacrylamide gels. The 25-kDa band from DEET-treated cells was enriched in peptides corresponding to glutathione S-transferase D10 and/or theta in the Aedes aegypti genome. Consistent with the increased expression of the labeled protein, DEET-treated cells had increased glutathione S-transferase activity, and the radiolabeled band bound to Sepharose 4B containing reduced glutathione. By analyzing partial tryptic digests, we established that DEET induces the homolog of A. aegypti glutathione S-transferase, class theta, corresponding to protein XP_001658009.1 in the NCBI database. This specific effect of DEET at the subcellular level suggests that DEET induces physiological responses that extend beyond recognition by the peripheral olfactory system.

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Culture of mosquito cells in eagle’s medium

Cited by 43 publications

References 9 publications

Sugar epitopes as potential universal disease transmission blocking targets

Sugar epitopes as potential universal disease transmission blocking targets

Cytological properties of an Aedes albopictus mosquito cell line infected with Wolbachia strain wAlbB

The insect repellent DEET (N,N-diethyl-3-methylbenzamide) increases the synthesis of glutathione S-transferase in cultured mosquito cells

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