Mild inactivation and inhibition of phenoloxidase in silkworm serum for xeno-free culture of insect cells
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- Hino Masato
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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- Karasaki Noriko
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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- Tatsuke Tsuneyuki
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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- Morokuma Daisuke
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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- Masuda Akitsu
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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- Mon Hiroaki
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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- Jae Man Lee
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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- Kusakabe Takahiro
- Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences
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抄録
<p> In insect cell culture, a fetal bovine serum is often used to maintain the cellular physiological conditions, irrespective of its relatively high cost and unstable supply. Several serum-free media for insects were developed, but it is challenging to adopt some insect cells to these serum-free media. Silkworm serum was used as a replacement of fetal bovine serum for a long time but not used widely due to the melanization of insect serum catalyzed by phenoloxidase (PO). PO inhibitors reported for insect serum preparation have a significantly negative effect on the long-term culture of healthy insect cells. In this study, we evaluated several kinds of PO inhibitors and found the method to prepare silkworm serum, which is suitable for the maintenance of insect cells and the production of the recombinant proteins by the baculovirus expression system. When L-Glutathione (reduced form) or L-Cysteine is used as a PO inhibitor at the time of recovery of silkworm serum, it is possible to suppress the browning of the silkworm serum. However, in this state of silkworm serum, browning is observed in long-term cell culture use. Therefore, it is preferable for cell culture to add 10% of silkworm serum treated at 50°C for 30 minutes to the medium with 2.7 mM L-Glutathione. Our results suggest that silkworm serum can be an alternative of mammalian serum for cell culture and used for producing the proteins for clinical application without any risk of contamination of zoonotic infectious disease viruses.</p>
収録刊行物
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- Journal of Insect Biotechnology and Sericology
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Journal of Insect Biotechnology and Sericology 89 (1), 1_009-1_016, 2020
社団法人 日本蚕糸学会
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詳細情報 詳細情報について
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- CRID
- 1390846609819343872
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- NII論文ID
- 130007821890
- 40022182079
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- NII書誌ID
- AA11558849
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- ISSN
- 18847978
- 13468073
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- NDL書誌ID
- 030302143
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- CiNii Articles
- KAKEN
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- 抄録ライセンスフラグ
- 使用不可