Cell Culture Fundamentals: Revival, Passaging, Cryopreservation

critical

Cell Death from Bacterial or Fungal Contamination

Media becomes rapidly turbid with visible particles or cloudiness. Cells show signs of toxicity, detachment, and death. Microscopy reveals motile bacteria or fungal filaments. Note: mycoplasma rarely causes acute cell death.

💡 4 · ✓ 6

severe

Cell Death from Poor Quality or Toxic Reagents

Sudden cell death occurs immediately after introducing new lot of media, serum, trypsin, or buffer. Previously healthy cultures show rapid necrosis or apoptosis within 12-24 hours.

💡 5 · ✓ 6

severe

Bacterial Contamination with Rapid Turbidity

Rapid-onset turbidity and color change of culture medium (when phenol red is present). Medium becomes cloudy and pH indicator changes color quickly.

💡 7 · ✓ 9

moderate

Contact Inhibition and Nutrient Depletion Cell Death

Cultures exceeding 80-100% confluence show central necrosis, detachment of cell sheets, and debris accumulation. Media becomes acidic (yellow) and depleted.

💡 4 · ✓ 5

moderate

Chemical Contamination from Residues

Poor cell growth, cell death, or altered cell behavior without visible microbial contamination. May include effects from free radicals, metal ions, disinfectant/detergent residues, or endotoxins persisting after bacterial contamination is cleared.

💡 6 · ✓ 6

moderate

Replicative Senescence After Excessive Passaging

Primary or finite cell lines show progressive slowing of growth, enlarged flattened morphology, failure to reach confluence, and increased spontaneous death after many passages.

💡 4 · ✓ 5