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第一株小菜蛾(Plutella xylostella)蛹发展起来的细胞系已经建成并命名为BCIRL-PX2-HNU3.BCIRL-PX2-HNU3是由20个剪碎的全蛹经过消化处理,培养在TC-199-MK 培养基中建立起来的.原代培养只历时48天就开始了第一次转代,由于细胞增殖快而稳定、此后即按1∶2的分裂比值,每周转代三次,待转代100次后,改按1∶10的比值转代,每周一次,至目前为止,该细胞已传至115代.BC1RL-PX2-HNU3细胞仅疏松贴附瓶壁,容易从瓶壁脱落.转代时,不需用消化酶.细胞的形状多种多样,但以园形和卵园形为主.28℃时,96小时细胞群体倍增时间为30小时±2.06.1×10~5/毫升细胞培养8天后,最大密度可达1.3×10~6细胞/毫升.该细胞系细胞的染色体粗短,多为多倍体,具有鳞翅目昆虫染色体的典型性状.用磷酸葡萄糖异构酶作等电聚焦,该细胞系与一些其他细胞系有明显差异.试验表明:BC1RL-PX2-HNU3细胞系对苜蓿尺蠖(Autographa californica)的核型多角体病毒以及中国棉铃虫(Heliothis armigera)病毒VHA—273敏感,并能复制出多角体.另外,用小菜蛾颗粒体病毒感染该细胞系,6天后,电镜切片证实培养的细胞胞质内出现了颗粒体病毒的蒴伏体,但在随后的多次试验中,未能重复.
The first line of Plutella xylostella pupae has been developed and named BCIRL-PX2-HNU3. BCIRL-PX2-HNU3 was digested with 20 cut whole pupae and cultured in TC-199- MK medium.The primary culture started the first round of transfection only 48 days after the first round of cell proliferation due to rapid and stable, then split ratio of 1: 2, three times a week, to be replaced by 100 Subsequent changes to a ratio of 1:10, once per week, so far this cell has been passed to 115. BC1RL-PX2-HNU3 cells loosely attached to the bottle wall, easy to fall off from the bottle wall. Digestive enzymes are not required.The shape of the cells is varied, but mainly in the shape of a park or egg.Under the conditions of 28 ° C, the cell population doubling time at 96 hours is 30 hours ± 2.06.1 × 10 -5 / ml cells After 8 days of cultivation, the maximum density can reach 1.3 × 10 ~ 6 cells / ml.The cells of the cell line are stubby and polyploid, with the typical characters of lepidopteran insect chromosomes. Electrofocusing, the cell line with some other cell lines were significantly different.The experiment showed that: BC1RL-PX2-HNU3 cell line alfalfa 蠖 (Autographa califo rnica), and the Chinese Heliothis armigera virus VHA-273, and could replicate polyhedra.In addition, the cell line was infected with Plutella xylostella virus, and after 6 days, it was confirmed by electron microscopy that cultured Granuloviruses were present in the cytoplasm of the cells, but were not repeated in subsequent trials.