Testing Hybridoma Cells for Mycoplasma Contamination

Area of Science:

• Microbiology
• Cell Biology
• Biotechnology

Background:

• Mycoplasmas are diverse microorganisms that frequently contaminate cell cultures, particularly hybridoma and myeloma lines.
• The difficulty in culturing mycoplasmas necessitates varied detection strategies.
• No single assay can identify all potential mycoplasma contaminants.

Purpose of the Study:

• To review and describe commonly used methods for detecting mycoplasma contamination in cell cultures.
• To highlight the importance of employing multiple detection techniques for comprehensive screening.

Main Methods:

• Mycoplasma screening via microbial growth on agar plates or in broth culture, under both aerobic and anaerobic conditions.
• Detection using the DNA-intercalating dye Hoechst 33258, which reveals bright, punctate cytoplasmic staining in contaminated cells.
• Utilization of commercial kits employing colorimetric assays or reporter cells for mycoplasma detection.

Main Results:

• Mycoplasma colonies exhibit a characteristic 'fried egg' appearance on agar plates, visible with an inverted microscope.
• Hoechst 33258 staining provides a rapid method for identifying mycoplasma DNA within fixed cells.
• Commercial kits offer alternative, often quicker, diagnostic options.

Conclusions:

• A combination of detection methods is recommended for thorough mycoplasma screening in cell cultures.
• Understanding the characteristics of different mycoplasma strains informs the choice of appropriate detection techniques.
• Effective mycoplasma detection is crucial for maintaining the integrity of hybridoma and myeloma cultures.