Related Experiment Video
Updated: Jun 28, 2026
Maintaining Wolbachia in Cell-free Medium
Published on: July 4, 2007
Maintaining Wolbachia in cell-free medium
Courtney Gamston1, Jason Rasgon
1Malaria Research Institute, Bloomberg School of Public Health, John Hopkins University, USA.
Abstract:
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.
Insights
This video protocol details purifying and maintaining viable Wolbachia bacteria outside of host cells. These extracellular Wolbachia are suitable for various downstream applications like micro-injection and DNA extraction.
Area of Science:
- Microbiology
- Cell Biology
- Parasitology
Background:
- Wolbachia are essential bacterial symbionts found in many arthropods.
- Maintaining Wolbachia viability outside of host cells is crucial for research applications.
Purpose of the Study:
- To provide a detailed video protocol for the purification and maintenance of extracellular Wolbachia.
- To demonstrate methods for assessing Wolbachia viability and suitability for downstream applications.
Main Methods:
- Purification of Wolbachia from cultured mosquito cells.
- Fluorescent viability assay for extracellular Wolbachia.
- Maintenance of purified Wolbachia in cell-free medium.
Main Results:
- Purified Wolbachia remain viable in cell-free medium for at least one week at room temperature.
- Extracellular Wolbachia do not replicate until re-inoculated into eukaryotic cells.
- Purified Wolbachia are suitable for micro-injection and DNA extraction.
Conclusions:
- This protocol enables the successful isolation and short-term preservation of viable extracellular Wolbachia.
- The method facilitates the use of Wolbachia in various experimental procedures, advancing research in host-symbiont interactions.
