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In vitro culture of polyembryonic wasp embryos

多胚黄蜂胚胎的体外培养

基本信息

批准号：
02660046
负责人：
IWABUCHI Kikuo
金额：
$ 1.41万
依托单位：
Tokyo University of Agriculture and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02660046/
关键词：
Polyembryony Parasitoid Embryo culture

项目摘要

Media suitable for culturing the 2-cell embryos of a polyembryonic wasp, Litomastix maculata, were exploited on more than 50 modified MGM-450 media. Bovine plasma albumin levels of more than I % was required as a component of the medium for the embryos to develop up to the complete morula. Ovalbum in and conalbumin also supported some development of the embryos, whereas lactalbumin was rather detrimental. The modifications of MGM-450 medium by changing the carbohydrates and the minerals to 0.2 5 % suct-ose alone and D-20's minerals, respectively, was required for the embryos to develop into Polygerms beyond the morula stac, . e. Experiments on the effect of supplementing- : the medium with insect hormones indicated that exogenous juvenile hormone(JH)supported the development of embryos into polygerms, which was a developmental- event specific to the polyembryonic insects. JH I was the most e erective !iorinone followed by 'H 11 and 'H Ill. On the other hand, 20-hydroxyecdysone did not. The composition of amino acids also was important for culturing 2-cell embryos up to polygerms ; amino acid composition of MGM-450 medium was the most suitable among those of several insect cell culture media. Supplementation of fetal bovine serum into the medium caused the higher percentage of Polygerm formation.There were not clear effects of albumin and juvenile hormone where the culture was started at the morula stage ; 80-90 % of morulae cultured in either MGM-450 medium or modified MGM-450 medium mentioned above developed into polygermis.

在50多种改良的MGM-450培养基上进行了适合培养多胚黄蜂（Litomastix maculata） 2细胞胚胎的培养基开发。牛血浆白蛋白水平必须超过1%，作为胚胎发育到完整桑葚胚的培养基成分。卵清蛋白和乳清蛋白对胚胎的发育也有一定的促进作用，而乳清蛋白则对胚胎发育有一定的不利影响。通过对MGM-450培养基进行改造，将碳水化合物和矿物质分别转化为0.2%的蔗糖和D-20的矿物质，可以使胚发育成超过桑葚胚的多胚。e.在培养基中添加昆虫激素的效果实验表明，外源幼虫激素（JH）支持胚胎发育成多胚，这是多胚昆虫特有的发育事件。JH：我是最有说服力的！iorinone后面是h11和hill。另一方面，20-羟基蜕皮激素则没有。氨基酸的组成对2细胞胚到多芽的培养也很重要；在几种昆虫细胞培养基中，MGM-450培养基的氨基酸组成最合适。在培养基中添加胎牛血清可提高多胚形成率。在桑葚胚期开始培养时，白蛋白和幼体激素对桑葚胚的影响不明显；在上述MGM-450培养基或改性MGM-450培养基中培养的桑葚胚80% - 90%发育成多芽。