Maintaining Wolbachia in Cell-free Medium


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This video describes a method for purifying Wolbachia pipientis from an Anopheles gambiae cell line and then culturing the endosymbiont in cell-free medium. An assay for viability of the bacterium is demonstrated.

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Gamston, C., Rasgon, J. Maintaining Wolbachia in Cell-free Medium. J. Vis. Exp. (5), e223, doi:10.3791/223 (2007).


In this video protocol, procedures are demonstrated to (1) purify Wolbachia symbionts out of cultured mosquito cells, (2) use a fluorescent assay to ascertain the viability of the purified Wolbachia and (3) maintain the now extracellular Wolbachia in cell-free medium. Purified Wolbachia remain alive in the extracellular phase but do not replicate until re-inoculated into eukaryotic cells. Extracellular Wolbachia purified in this manner will remain viable for at least a week at room temperature, and possibly longer. Purified Wolbachia are suitable for micro-injection, DNA extraction and other applications.


The authors have nothing to disclose.


1 Comment

  1. Dear sir
    I am interest of your work, Wolbachia, so i have some question that make me clear about culture bacteria endosymbionts as
    1. How did you infected Wolbachia in your medium?
    ². What's the strain of your laboratory wolbachia?
    Please forward to me if possible and i woud like to thank for your kind

    Best reguard

    Veerayuth Kittichai
    Entomology Unit
    Chulalongkorn University

    Posted by: Anonymous
    September 15, 2010 - 10:00 PM

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