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Cell Culture (Contamination) critical

Mycoplasma Contamination - Silent Growth Inhibitor

Symptom
Slowed cell growth, altered cellular metabolism, chromosomal aberrations, and interference with cell attachment. No visible turbidity in medium. Difficult to detect by light microscopy due to small size (0.15-0.3 µm).
Common Causes
  1. 1 Introduction through poor aseptic technique (mycoplasma found on human skin)
  2. 2 Contaminated fetal bovine serum and other supplements
  3. 3 Highly transmissible among neighboring contaminated cultures
  4. 4 Standard 0.22 µm or 0.45 µm filtration fails to exclude mycoplasma
  5. 5 Unaffected by cell wall-targeting antibiotics due to lack of cell walls
  6. 6 Mycoplasma titer can reach 10^8 organisms/mL without causing visible turbidity
Solutions
  1. 1 Establish routine mycoplasma screening using culture, DNA staining (DAPI/Hoechst), or PCR-based detection
  2. 2 Use filters with 0.1 µm or smaller pores for media and buffer filtration
  3. 3 Implement strict adherence to good laboratory practices
  4. 4 Test all incoming cell lines in quarantine for two weeks without antibiotics
  5. 5 Use mycoplasma detection and elimination kits from reputable suppliers
  6. 6 Discard contaminated cultures immediately to prevent lab-wide spread
  7. 7 Avoid routine antibiotic use which masks contamination and creates resistant strains
Related Video (2)
Bilibili (China-Accessible Mirrors) ★ 85
Complete Cell Culture Protocol: Revival, Passaging, Cryopreservation
"Explicitly covers contamination recognition and critical precautions alongside passaging, directly addressing prevention of the mycoplasma introduction pathway through aseptic technique."
Bilibili (China-Accessible Mirrors) ★ 82
Cell Culture Training Video — Aseptic Technique & Routine Maintenance
"Dedicated focus on aseptic technique in the biosafety cabinet and routine maintenance—the core prevention strategy for mycoplasma contamination from skin contact."
Source: sigmaaldrich.com ↗
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