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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 (3)
Bilibili (China-Accessible Mirrors) ★ 88
Cell Culture Training Video — Aseptic Technique & Routine Maintenance
"Focuses specifically on aseptic technique in the biosafety cabinet and troubleshooting, directly addressing the contamination source."
Bilibili (China-Accessible Mirrors) ★ 78
Complete Cell Culture Protocol: Revival, Passaging, Cryopreservation
"Comprehensive hands-on protocol that explicitly includes contamination recognition and critical precautions alongside cell culture fundamentals."
Bilibili (China-Accessible Mirrors) ★ 72
Cell Passaging Steps and Precautions
"Passaging demonstration with explicit attention to contamination prevention, showing the correct technique whose failure leads to mycoplasma introduction."
Source: sigmaaldrich.com ↗
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