一种体外分析亚洲玉米螟幼虫血细胞包囊的改进方法

An Improved Method on Encapsulation Assays by Larval Hemocytes from Ostrinia furnacalis in vitro

包囊反应是昆虫清除侵入体内的外源物如病原物和寄生生物的一种非常重要的细胞免疫反应。由于受到不便于观察、操作复杂等问题的限制,很多包囊分析实验无法或难于在昆虫体内完成。体外包囊方法在一定程度上解决了这些问题。目前常用的体外包囊是在96孔板中加入昆虫血细胞和外源物,如凝胶珠进行培养观察,但这种方法存在着一些明显的缺陷。本文以亚洲玉米螟(Ostrinia furnacalis)幼虫血细胞为研究对象,使用0.2mL的离心管(Eppendorf tube)代替96孔板,并将其固定在匀速旋转的载体上培养,极大程度的模拟了反应物在昆虫体内的状态。结果表明,改进方法后昆虫血细胞的体外包囊效果得到明显提高,且血细胞的状态也得到明显改善;而添加抗凝剂会减弱血细胞对外源物的包囊能力。

Encapsulation, a kind of cellular immune reaction of insect, plays an important role in eliminating foreign objects such as pathogens and parasites which invade insect hemocoels. As a result of limited observation and complicated manipulations, analysis of encapsulation can not be easily performed in vivo. Encapsulation in vitro resolves these problems to a certain extent. Now, encapsulation assays in vitro are usually performed in 96-well plates in which foreign objects such as sephadex beads and insect hemocytes are contained and observed, but some shortcomings exist in this method. In this paper, an improved analysis method on the encapsulation of Ostriniafurnacalis larvae hemocytes in vitro is introduced. 0.2 mL Eppendorf tubes are the substitute for the 96-well plates and they are fixed to a revolving rotator at a speed of 4 circles per min. The states of hemocytes and sephadex beads in insect＇s hemocoels are replicated under this method. Results showed that the states of hemocytes were improved and encapsulation was enhanced under the new method and anticoagulants might inhibit the encapsulation ability of hemocytes.