USDA, ARS, US Horticultural Research Laboratory, 2001 South Rock Road, Ft. Pierce, FL 34945, USA.
In Vitro Cell Dev Biol Anim. 2010 Feb;46(2):83-6. doi: 10.1007/s11626-009-9246-x. Epub 2009 Dec 24.
A media for the production of cell cultures from hymenopteran species such as honey bee, Apis mellifera L. (Hymenoptera: Apidae) was developed. Multiple bee cell cultures were produced when using bee larvae and pupae as starting material and modified Hert-Hunter 70 media. Cell culture systems for bees solves an impasse that has hindered efforts to isolate and screen pathogens which may be influencing or causing colony collapse disorder of bees. Multiple life stages of maturing larvae to early pupae were used to successfully establish cell cultures from the tissues of the head, thorax, and abdomen. Multiple cell types were observed which included free-floating suspensions, fibroblast-like, and epithelia-like monolayers. The final culture medium, WH2, was originally developed for hemipterans, Asian citrus psyllid, Diaphorina citri, and leafhopper, Homalodisca vitripennis cell cultures but has been shown to work for a diverse range of insect species such as bees. Bee cell cultures had various doubling times at 21-23 degrees C ranging from 9-15 d. Deformed wing virus was detected in the primary explanted tissues, which tested negative by rt-PCR for Israeli acute paralysis virus (IAPV), Kashmir bee virus, acute bee paralysis virus, and black queen cell virus. Culture inoculation with IAPV from an isolate from Florida field samples, was detectable in cell cultures after two subcultures. Cell culture from hymenoptera species, such as bees, greatly advances the approaches available to the field of study on colony collapse disorders.
开发了一种用于生产膜翅目物种（如蜜蜂，Apis mellifera L.（膜翅目：Apidae））细胞培养物的培养基。使用蜜蜂幼虫和蛹作为起始材料，并对改良的 Hert-Hunter 70 培养基进行操作，可产生多种蜜蜂细胞培养物。蜜蜂细胞培养系统解决了一个僵局，该僵局阻碍了分离和筛选可能影响或导致蜜蜂种群崩溃紊乱的病原体的努力。成熟幼虫到早期蛹的多个生命阶段被用于成功地从头部、胸部和腹部组织中建立细胞培养物。观察到多种细胞类型，包括游离悬浮液、成纤维细胞样和上皮样单层。最终的培养基 WH2 最初是为半翅目昆虫、亚洲柑橘木虱 Diaphorina citri 和叶蝉 Homalodisca vitripennis 细胞培养开发的，但已证明它适用于多种昆虫物种，如蜜蜂。在 21-23 摄氏度下，蜜蜂细胞培养物的倍增时间从 9-15 天不等。在原代外植组织中检测到变形翅膀病毒，但通过 rt-PCR 检测以色列急性麻痹病毒（IAPV）、喀什米尔蜜蜂病毒、急性蜜蜂麻痹病毒和黑皇后细胞病毒均为阴性。从佛罗里达州田间样本分离株接种 IAPV 后，可在两次传代培养后在细胞培养物中检测到。膜翅目物种（如蜜蜂）的细胞培养极大地推进了对蜂群崩溃紊乱研究领域的研究方法。
