Isolation and primary culture of stem cells from the midgut epithelium of the silkworm Bombyx mori
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- Fujiwara Nozomi
- The United Graduate School of Agricultural Sciences, Tottori University
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- Azuma Masaaki
- The United Graduate School of Agricultural Sciences, Tottori University
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- Itoh Masanobu
- Kyoto Institute of Technology
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- Kobayashi Jun
- Graduate School of Sciences and Technology for Innovation, Yamaguchi University
書誌事項
- タイトル別名
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- Isolation and primary culture of stem cells from the midgut epithelium of the silkworm <i>Bombyx mori</i>
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抄録
<p> The process of molting is essential for the growth of lepidopteran larvae and their metamorphosis into adults. However, the investigation of cellular renewal and regeneration is limited by the lack of efficient methods for the isolation and characterisation of midgut stem cells in vitro. Here, we demonstrate an easy and secure protocol for midgut stem cell isolation from molting larvae and pharate pupae of the silkworm, Bombyx mori. This meth- od is based on the physiology of stem cells, which lie loosely associated with the base of epithelial cells and multiply during the molting or metamorphosing phase. They finally intercalate or replace existing epithelial cells as they elongate and differentiate into mature forms. Using approximately 10 insects, it was possible to obtain >500,000 cells with high homogeneity and >80% viability. Stem cells prepared from the fourth instar molting larvae and pharate pupae were spherical and could be preserved at 4°C for 1-2 weeks with no viability loss. However, the number of living cells showed a steady decline when cultured in Grace’s insect culture medium for one week. Supplements such as fetal bovine serum, lipid-rich bovine serum albumin, or heat-treated native B. mori plasma did not support stem cell proliferation or suppress cell death. Although continuous stem cell culture is still difficult, the described method for midgut investigation could potentially utilized for biochemical studies of stem cells and pathological assays in vitro, which may lead to novel strategies for agricultural pest management.</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 (3), 3_055-3_062, 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
- 1390287142238993792
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- NII論文ID
- 130007987477
- 40022471184
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- NII書誌ID
- AA11558849
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- ISSN
- 18847978
- 13468073
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- NDL書誌ID
- 031261972
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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