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Development and Application of Insect Cell Culture Techniques

Research Project

Project/Area Number 01860008
Research Category

Grant-in-Aid for Developmental Scientific Research (B).

Allocation TypeSingle-year Grants
Research Field 蚕糸学
Research InstitutionKyushu University

Principal Investigator

KAWARABATA Takeshi  Professor, Faculty of Agriculture, Kyushu University, 農学部, 教授 (60038221)

Co-Investigator(Kenkyū-buntansha) NAGATA Masao  Associate Professor, Faculty of Agriculture, Tokyo University, 農学部, 助教授 (70107407)
KOBAYASHI Michihiro  Associate Professor, Faculty of Agriculture, Nagoya University, 農学部, 助教授 (60111837)
OHBA Michio  Associate Professor, Faculty of Agriculture, Kyushu University, 農学部, 助教授 (80038281)
KOBAYSHI Jyun  Research Scientist, Genetics and Breeding Division, National Institute of Sericu, 蚕糸・昆虫農業技術研究所・遺伝育種部, 研究員
MITSUHASHI Jyun  Professor, Faculty of Agriculture, Tokyo University of Agriculture and Technolog, 農学部, 教授 (90209809)
井上 元  農林水産省, 蚕糸・昆虫農業技術研究所・遺伝育種部, 室長
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 1990: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1989: ¥7,700,000 (Direct Cost: ¥7,700,000)
KeywordsCulture of microsporidians / Low cost media (MM) / Medium MGM-450 / Colonial coloning / Assay of bacterial toxins / Hemolymph factor (s) / Serological identification / カイコBMーN細胞系 / 鱗翅目昆虫病原性微胞子虫 / 鱗翅目昆虫由来培養細胞 / 定着性昆虫培養細胞 / 浮遊性昆虫培養細胞 / 低コスト昆虫細胞用培地 / カイコ幼虫体液添加 / 昆虫ウイルス産生促進効果 / 抗BMーN細胞ウサギ血清
Research Abstract

Infection and development of microsporidians (Nosema bombycis NIS 001, N. sp. NIS M11, N. sp. F-30, N. mesnili AH, Thelohania sp. K-25) pathogenic to lepidopterous insects were studied in a cell line of Antheraea eucalypti. Microsporidian spores primed with 0.1 N potassium hydroxide solution were mixed with the A. eucalypti cell cultures and infected cultures were maintained for more than 18 months.
Low cost insect cell culture media (MM) and its application to spinner culture of insect cells were investigated. Growth of two lepidopteran and one dipteran cell lines was compared under still and spinner culture conditions. Apparently, the insect cells cultured in plastic culture flasks showed higher growth rates when smaller culture vessels were employed.
Also, a general purpose culture medium (MGM-450) for insect (Lepidopera, Diptera, Hemiptera and Coleoptera) cells was newly formulated.
Cloning of insect cells using Bombyx mori cell line (SES-BoMo-15AII) was studied and establishment of the B. mori cell line resulted from a single cell was unsuccessful. It was suggested that a colonial coloning was practical method for cloning of insect cells at the present time.
Spodoptera furgiperda Sf21AEII cells were highly sensitive to heat labile cytopathic factor (s) produced by several serovars of Bacillus thuringiensis. However, in vitro assay of the heat stable exotoxin (fly toxin) of B. thuringiensis using the similar system showed negative results.
Addition of insect hemolymph factor (s) to Bombyx mori BM-N cells infected with a nuclear polyhedrosis virus, markedly enhanced the production of viral structural and inclusion proteins in vitro.
Specific and common cell surface antigens of cultured lepidopteran cells were demonstrated and serological identification of insect cell lines was applicable for several insect cell lines.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Takeshi Kawarabata: "Infection and development of Thelohania sp.Kー25(Microsporida:Thelohaniidae)in lepidopteran cell lines." Applied Entomology and Zoology. 26. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 三橋 淳: "昆虫細胞の浮遊攪拌培養" 日本応用動物昆虫学会誌. 35. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 小林 淳・井上 元・今西 重雄: "カンボ-ジュ卵巣から樹立された培養細胞系とそのウイルス感受性" 日本蚕糸学雑誌. 60. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Michio Ohba and Hidenori Iwahana: "Intrahemocoelic virulence of Bacillus thuringiensis correlates with in vitro insectーcell killing activity in bacterial culture supernatant" Letters in Applied Microbiology. 12. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 永田 昌男: "カイコ由來培養細胞表面抗原の酵素免疫測定法による解析" 日本蚕糸学雑誌. 60. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Michihiro Kobayashi: "Protein synthesis in BMーN cells infected with Bombyx mori nuclear polyhedrosis virus." Journal of Invertebrate Pathology. 57. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Takeshi Kawarabata: "Infection and development of Thelohania sp. K-25 (Microsporida : Thelohaniidae) in a lepidopteran cell line." Applied Entomology and Zoology. 26. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Jyun Mitsuhashi: "Suspension culture of lepidopteran and dipteran cell lines. (In Japanese)" Japanese Journal of Applied Entomology and Zoology. 35. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Jyun Kobayashi, Hajime Inoue, and Sigeo Imanishi: "Establishment and viral susceptibility of a Bombyx mori cell line from ovarian tissues of Cambodge strain. (In Japanese)" Journal of Sericultural Science of Japan. 60. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Michio Ohba and Hidenori Iwahana: "Intrahemocoelicvirulence of Bacillus thuringiensis correlates with in vitro insect-cell killing activity in bacterial culture supernatant." Letters in Applied Microbiology. 12. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Masao Nagata: "Serological comparison of cell surface antigens in lepipdopteran cell lines. (In Japanese)" Journal of Sericultural Science of Japan. 60. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Michihiro Kobayashi: "Protein synthesis in BM-N cells infected with Bombyx mori nuclear polyhedrosis virus." Journal of Invertebrate Pathology. 57. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Takeshi Kawarabata: "Infection and development of <Thelohania>___ー sp.Kー25 (Microsporida:Thelohaniidae) in lepidopteran cell lines." Applied Entomology and Zoology. 26. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 三橋 淳: "昆虫細胞の浮遊撹拌培養" 応用動物昆虫学雑誌. 35. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 小林 淳・井上 元・今西 重雄: "カンボ-ジュ卵巣から樹立された培養細胞系とそのウイルス感受性" 日本蚕糸学雑誌. 60. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Michio Ohba and Hidenori Iwahana: "Intrahemocoelic virulence of <Bacillus>___ー <thuringiensis>___ー correlates with in vitro insectーcell killing activity in bacterial culture supernatant" Letters in Applied Microbiology. 12. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 永田 昌男: "カイコ由来培養細胞表面抗原の酵素免疫測定法による解析" 日本蚕糸学雑誌. 60. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Michihiro Kobayashi: "Protein synthesis in BMーN cells infected with <Bombyx>___ー <mori>___ー nuclear polyhedrosis virus." Journal of Invertebrate Pathology. 57. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Takeshi Kawarabata: "Development of Nosema mesnili AH-1(Microsporida:Nosematidae)causing acute paralysis on the larvae of Spodoptera litura(Lepidoptera:Noctuidae)in insect cell cultures" Applied Entomology and Zoology. 25. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Takeshi Kawarabata: "Infection and development of Thelohania sp.K25(Microsporida:Thelohaniidae)isolated from an adult of the common cabbageworm,Pieris rapae curcivora in lepidopteran cell lines" Applied Entomology and Zoology. 25. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Takeshi Kawarabata: "Factors affecting the sporogenesis of Thelohania sp.K25(Microsporida:Thelohaniidae)in an insect cell line of Antheraea eucalypti" 日本蚕糸学雑誌. 59. (1990)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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