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Cell-assisted culture of fastidious spiroplasmas: initial analysis of growth factors
K J Hackett1, D E Lynn, A S Ginsberg
1Plant Protection Institute, U.S. Department of Agriculture, Beltsville, MD 20705.
Abstract:
Half of the spiroplasmas observed microscopically in insects cannot be cultivated and are thus inaccessible to study. Media mixed with cultured insect cells have now been used to isolate two of these spiroplasmas--the sex-ratio organism (SRO) of Drosophila and the Colorado potato beetle spiroplasma (CPBS). Studies described herein indicate that at least one of the cell-supplied factors is involved in redox maintenance. A wide variety of insect cell culture systems were suitable for primary isolation of the CPBS. The SRO and CPBS were found to attach to insect cells in vitro.
Insights
Researchers developed a new method using insect cell cultures to study unculturable spiroplasmas. This breakthrough allows investigation of important insect pathogens like the sex-ratio organism (SRO) and Colorado potato beetle spiroplasma (CPBS).
Area of Science:
- Microbiology
- Insect Pathology
- Cell Biology
Background:
- Many spiroplasmas, important insect pathogens, are difficult to culture.
- This lack of cultivation limits research into their biology and impact.
Purpose of the Study:
- To develop a novel method for culturing unculturable spiroplasmas.
- To investigate the role of insect cells in spiroplasma isolation and survival.
Main Methods:
- Utilized insect cell culture media for spiroplasma isolation.
- Co-cultured spiroplasmas with various insect cell systems.
- Observed spiroplasma attachment to insect cells in vitro.
Main Results:
- Successfully isolated two previously unculturable spiroplasmas: Drosophila's sex-ratio organism (SRO) and Colorado potato beetle spiroplasma (CPBS).
- Identified cell-supplied factors involved in redox maintenance for spiroplasma survival.
- Demonstrated that SRO and CPBS attach to insect cells.
Conclusions:
- Insect cell co-culture is an effective method for isolating unculturable spiroplasmas.
- Insect cells provide essential factors, including those for redox maintenance, supporting spiroplasma growth.
- Spiroplasma attachment to host cells is a key interaction.