Serum-free culture of an embryonic cell line from Bombyx mori and reinforcement of susceptibility of a recombinant BmNPV by cooling
| Autor: | Sekine Toshiaki, Jun Kobayashi, Shigeo Imanishi |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
viruses
Genetic Vectors Cell Culture Techniques Gene Expression Culture Media Serum-Free Cell Line Bombycidae Bombyx mori Hemolymph Doubling time Animals biology fungi Temperature Nuclear Polyhedrosis Virus Embryo Cell Biology General Medicine biology.organism_classification Bombyx Virology Molecular biology Embryonic stem cell Nucleopolyhedroviruses Cell culture Developmental Biology |
| Zdroj: | In vitro cellulardevelopmental biology. Animal. 48(3) |
| ISSN: | 1543-706X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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