Evaluation of three methods for curing hybridomas from mycoplasma contamination

Area of Science:

• Cell Biology
• Microbiology
• Biotechnology

Background:

• Hybridomas, crucial for antibody production, are vulnerable to mycoplasma contamination.
• Mycoplasma contamination compromises hybridoma function and experimental reliability.

Purpose of the Study:

• To compare the efficacy of different methods for eradicating mycoplasma contamination in hybridomas.
• To identify the most efficient, cost-effective, and reliable decontamination strategy.

Main Methods:

• In vitro treatment using 5-bromouracil (5-BrUra) with photosensitization.
• In vitro treatment using BM-cycline.
• In vivo decontamination via passage through mice.

Main Results:

• BM-cycline demonstrated superior efficiency, requiring fewer treatments (six) compared to 5-BrUra (ten) for highly contaminated hybridomas.
• BM-cycline achieved a 50% reduction in contamination after the first treatment.
• Hybridomas treated with BM-cycline maintained stability and immunoglobulin secretion after 20 passages, unlike one clone treated with 5-BrUra which showed recurrence.

Conclusions:

• BM-cycline is the preferred method for hybridoma mycoplasma decontamination due to its higher efficiency, reduced treatment time, simplicity, and lower cost.
• Photosensitization and in vivo mouse passage were less consistently effective.
• Bovine-origin mycoplasma strains, including Mycoplasma arginini and Acholeplasma laidlawii, were identified as contaminants.