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Establishment and characterization of two new cell lines from the mosquito Armigeres subalbatus (Coquillett) (Diptera: Culicidae)

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Abstract

Armigeres subalbatus (Coquillett) is a medically important mosquito and a model species for immunology research. We successfully established two cell lines from the neonate larvae of A. subalbatus using two different media. To our knowledge, this is the first report of an established Armigeres mosquito cell line. The cell lines, designated as Ar-3 and Ar-13, consist of adherent and diploid cells with compact colonies. Both these cell lines grow slowly after passage at a split ratio of 1:5 and a population doubling time of 2.7 and 3.0 d, respectively. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to confirm that these lines correspond to the species of origin and are clearly distinct from seven other insect cell lines. Furthermore, reverse-transcription PCR was used to demonstrate that the Ar-3 cell line is susceptible to the Japanese encephalitis virus and two insect flaviviruses associated with Culex and Aedes mosquitoes but relatively insensitive to dengue virus. These data indicate that the newly established cell lines are cellular models of A. subalbatus as well as beneficial tools for the propagation of viruses associated with the Armigeres mosquito.

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Acknowledgments

This work was supported by the Japan Society for the Promotion of Science, Grant Number 25870210 to KH, and Grant Number 25305010 to HI, KS, and TS. This work was also supported in part by grants-in-aid of the Japanese Ministry of Health, Labor and Welfare (H24-Shinko-Ippan-007).

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    Authors and Affiliations

    1. Department of Medical Entomology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan

      Keita Hoshino, Haruhiko Isawa, Ryusei Kuwata, Kyoko Sawabe, Mutsuo Kobayashi & Toshinori Sasaki

    2. Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai, Fuchu, Tokyo, 183-8509, Japan

      Keita Hoshino & Kikuo Iwabuchi

    3. Department of Virology 1, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan

      Shigeru Tajima & Tomohiko Takasaki

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    1. Keita Hoshino
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    2. Haruhiko Isawa
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    3. Ryusei Kuwata
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    4. Shigeru Tajima
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    5. Tomohiko Takasaki
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    6. Kikuo Iwabuchi
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    8. Mutsuo Kobayashi
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    9. Toshinori Sasaki
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    Correspondence to Toshinori Sasaki.

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    Editor: T. Okamoto

    Keita Hoshino and Haruhiko Isawa contributed equally to this work.

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    Hoshino, K., Isawa, H., Kuwata, R. et al. Establishment and characterization of two new cell lines from the mosquito Armigeres subalbatus (Coquillett) (Diptera: Culicidae). In Vitro Cell.Dev.Biol.-Animal 51, 672–679 (2015). https://doi.org/10.1007/s11626-015-9883-1

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