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Serum-free culture of an embryonic cell line from Bombyx mori and reinforcement of susceptibility of a recombinant BmNPV by cooling

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Abstract

We established the first continuous cell line that uses a serum-free culture from the embryo of Bombyx mori (Lepidoptera: Bombycidae), designated as NIAS-Bm-Ke17. This cell line was serially subcultured in the SH-Ke-117 medium. The cells adhere weakly to the culture flask, and most cells have an oval shape. The cell line was subcultured 154 times, and the population doubling time is 83.67 ± 5.22 h. Random amplification of polymorphic DNA-polymerase chain reaction with a tenmar single primer for discrimination of insect cell lines recognized the NIAS-Bm-Ke1 cell line as B. mori. This cell line does not support the growth of the B. mori nuclear polyhedrosis virus (BmNPV) in the absence of the heat-inactivated hemolymph of B. mori. However, the heat-inactivated hemolymph in 1% volume of the medium supported a high level of susceptibility to BmNPV. In addition, the cooling treatment of the cells at 2.5°C also enhanced the susceptibility. We report a new serum-free culture system of the B. mori cell line for the baculovirus expression vector system.

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Acknowledgements

We thank Dr. S. Tomita (Transgenic Silkworm Research Unit, National Institute of Agrobiological Sciences) for his kind advice. This work was supported by grants for research from the Research and Development Program for New Bio-industry Initiatives.

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Authors and Affiliations

  1. Department of Insect Science, National Institute of Agrobiological Sciences, 1-2, Owashi, Tsukuba, Ibaraki, 305-8634, Japan

    Shigeo Imanishi

  2. Laboratory Agrobiology, Department of Biological and Environmental Sciences, Faculty of Agriculture, Yamaguchi University, 1677-1, Yoshida, Yamaguchi, 753-8525, Japan

    Jun Kobayashi

  3. SHIMA Laboratories Co., Ltd. 3-6-10, Maeno-cho, Itabashi-ku, Tokyo, 174-0063, Japan

    Toshiaki Sekine

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  1. Shigeo Imanishi
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  2. Jun Kobayashi
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  3. Toshiaki Sekine
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Correspondence to Shigeo Imanishi.

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Editor: T. Okamoto

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Imanishi, S., Kobayashi, J. & Sekine, T. Serum-free culture of an embryonic cell line from Bombyx mori and reinforcement of susceptibility of a recombinant BmNPV by cooling. In Vitro Cell.Dev.Biol.-Animal 48, 137–142 (2012). https://doi.org/10.1007/s11626-011-9465-9

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