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Development and Application of Insect Cell Culture Techniques

昆虫细胞培养技术的开发与应用

基本信息

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

项目摘要

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.

家蚕微孢子虫Nosema bombycis NIS 001、N. nisM 11，新脉轮藻N. sp. F-30，N. mesnili AH，Thelohania sp. K-25）对鳞翅目昆虫的致病性。将用0.1N氢氧化钾溶液引发的微孢子虫孢子与A.本实验研究了低成本昆虫细胞培养基（MM）及其在昆虫细胞转瓶培养中的应用。在静止和旋转培养条件下比较了两种鳞翅目和一种双翅目细胞系的生长。本研究还配制了一种昆虫细胞通用培养基（MGM-450），并利用家蚕细胞系（SES-BoMo-15 AII）进行了昆虫细胞克隆的研究，建立了昆虫细胞克隆的B.从单个细胞产生的Mori细胞系是不成功的。结果表明，集落克隆法是目前昆虫细胞克隆的一种实用方法，且对苏云金芽孢杆菌（Bacillus thuringiensis）的几个血清型产生的不耐热细胞病变因子高度敏感。然而，在体外测定B的热稳定外毒素（蝇毒素）。在家蚕BM-N细胞中加入昆虫血淋巴因子，能显著提高病毒结构蛋白和包涵体蛋白的产量，证明了鳞翅目昆虫细胞表面的特异性和共同性抗原，并证明了昆虫细胞系的血清学鉴定适用于几种昆虫细胞系。