Submitted to: In Vitro Cellular and Developmental Biology - Animals
Publication Type: Peer reviewed journal
Publication Acceptance Date: 9/29/2009
Publication Date: 1/2/2010
Citation: Hunter, W.B. 2010. Medium for development of bee cell cultures (Apis mellifera: Hymenoptera: Apidae). In Vitro Cellular and Developmental Biology - Animals. 46:83-86. Interpretive Summary: A honey bee cell culture system was developed using larvae and pupae of the honey bee, Apis mellifera. This culture system solves a critical roadblock to bee research and provide a unique means to study virus-bee cell interactions. The lack of cell cultures from hymenoptera (bees, wasps, ants) has prevented the advance of many research projects. Bee cell cultures will now provide an essential tool that has been lacking in apiculture research to better understand bee diseases and address Colony Collapse Disorder. The susceptible nature of honey bees to virus infections includes over 17 different viruses. The established cell lines will further assist in the independent isolation of each virus, thus setting up a means to evaluate independent viral effects or multiple viral effects leading to bee death. The importance of bees as pollinators and the recent emergence of Colony Collapse Disorder highlighted the need for a bee cell culture system. These cell lines will provide a means to isolate and propagate individual virus strains for further evaluation of the impact and role of Israeli Acute Paralysis virus, IAPV, and the other viruses known to infect bees and other insects.
Technical Abstract: A bee cell culture system was developed. A medium, WH2, 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 the 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, CCD, of bees. Multiple life stages, of maturing larvae to early pupae were used to successfully establish cell cultures from the head, thorax, and abdomen tissues. 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, 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'C ranging from 9-15 days. Deformed Wing Virus, DWV, was detected in the primary explanted tissues, which tested negative by rtPCR for Israeli Acute Paralysis Virus, Kashmir Bee Virus, KBV, Acute Bee Paralysis Virus, ABPV, and Black Queen Cell Virus, BQCV. 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.