Development and Application of Insect Cell Culture Techniques

昆虫细胞培养技术的开发与应用

• 批准号: 01860008
• 负责人: KAWARABATA Takeshi
• 金额: $ 7.49万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B).
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01860008/
• 关键词: Culture 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

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.

菌孢子虫的感染和发育（Nosema bombycis Nis 001，N。sp。NisM11，N。sp。F-30，N。MesniliAH，Thelohaniasp。K-25）在鳞翅目Eucalypti的细胞系中研究了对鳞翅目昆虫的致病性。将氢氧化钾溶液引发的微孢子孢子与桉树曲霉培养物混合，并将感染的培养物维持了18个月以上。低成本的昆虫细胞培养基（MM），并研究了其在昆虫细胞的旋转培养物中的应用。在静止和旋转器培养条件下，比较了两个鳞翅目和一只双翅目细胞系的生长。 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单细胞没有成功。有人提出，殖民地结肠是目前克隆昆虫细胞的实用方法。Spodopterafurgiperda sf21aeii细胞对硫代素芽孢杆菌的几种血清co产生的热不稳定细胞病变因子（S）高度敏感。然而，使用相似系统的热稳定外毒素（蝇毒素）的体外测定显示出阴性结果。对昆虫血淋巴因子（S）的影响（S）的影响（S）被核多刺激病毒感染了Bombyx Mori BM-N细胞，显着增强了病毒结构和含有蛋白质的植物含量的生产。昆虫细胞系的血清学鉴定适用于几种昆虫细胞系。

项目成果

Takeshi Kawarabata: "Infection and development of Thelohania sp. K-25 (Microsporida : Thelohaniidae) in a lepidopteran cell line." Applied Entomology and Zoology. 26. (1991)

Takeshi Kawarabata：“Thelohania sp. K-25（微孢子虫：Thelohaniidae）在鳞翅目细胞系中的感染和发育。”

Takeshi Kawarabata: "Infection and development of <Thelohania>___ー sp.Kー25 (Microsporida:Thelohaniidae) in lepidopteran cell lines." Applied Entomology and Zoology. 26. (1991)

Takeshi Kawarabata：“鳞翅目细胞系中 <Thelohania>___ sp.K-25（微孢子虫：Thelohaniidae）的感染和发育。”应用昆虫学和动物学 26。

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)

Jyun Kobayashi、Hajime Inoue 和 Sigeo Imanishi：“来自 Cambodge 品系卵巢组织的家蚕细胞系的建立和病毒敏感性。（日语）”日本蚕业科学杂志。

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)

Masao Nagata：“酶免疫分析培养蚕细胞表面抗原”，日本血清学杂志 60。（1991）

Michihiro Kobayashi: "Protein synthesis in BMーN cells infected with <Bombyx>___ー <mori>___ー nuclear polyhedrosis virus." Journal of Invertebrate Pathology. 57. (1991)

Michihiro Kobayashi：“感染 <Bombyx>___- <mori>___- 核多角体病毒的 BM-N 细胞中的蛋白质合成。无脊椎动物病理学杂志 57。”