甜菜夜蛾幼虫脂肪体细胞建系方法的探讨

Establishment of Cell Lines from the Fat Bodies of Spodopter exigua Larvae

本研究以甜菜夜蛾幼虫脂肪体组织为材料,探讨最初该细胞通过原代培养从组织中游离出不断增殖,经过数次传代,最终建立细胞系的过程以及培养过程中污染控制、培养基结晶对原代培养的影响等。结果表明,贴壁处理对原代细胞的游离具有重要作用,经贴壁处理,有62%的培养出现游离细胞;游离出来的细胞持续增殖,能够布满培养表面并进行传代的比率为33%;最终有15%的原代培养建系成功。起始培养至首次传代成功的平均时间是85天。污染和培养基结晶是原代培养失败的重要原因之一。

This study focused on how to initiate insect cell lines from primary culture using the fat bodies of Spodoptera exigua larvae as culture tissue. Factors influencing proliferation of new cells from the primary culture, how to deal with subculture, as well as impact of microorganism pollution or medium crystallization on the primary culture were investigated and discussed. The results showed that the treatment that ensured the cultured tissue fast adhere to the culture surface was very important for new cells proliferating from the tissue. In the all adhesion cultures there were 62% of the cultures abserved cell proliferation. Those continuous proliferating and further covering the whole culture surface were of 33%. Finally, approximate 15% of the primary cultures were developed to continuous cell lines. The average time from initial training to the success of the first passage was 85 days. Pollution and crystallization of the primary culture were important causations of failure.