Cell culture contamination seminar

Tuesday, June 13, 2017 10:16 AM

Types of contamination
Virus
no visible sign
Make sure BSC is working
Bacteria and fungi
Most common
Cloudy or turbid medium
Furry clumps
Very quickly cause cell death by pH shifts and taking up nutrients
Mycoplasma
Most serious, widespread
Not detectable with regular microscopy, need to test to find (DNA stain)
Technically bacteria but much smaller
No cell walls -> Resistant to most antibiotics
Can sometimes slip through filter caps
Affect host cell's metabolism, morphology
Cell growth slow, transfection efficiency low, cells may clump
Cross contamination
With other mammalian cell types
Occurs when the foreign cell type is better adapted to the culture conditions and
grows faster
Serious issue - only solution is to use cell lines confirmed by DNA fingerprinting
Volatile organic compounds (VOCs)
Commonly used lab volatile chemicals easily dissolve in culture media - acetone,
ether, phenol, etc
Cleaners - bleach
Increasing use of cell culture = increased risk
Contaminants from YOU
Contamination occurs when technique deteriorates
Work with only one cell type at a time, don't share reagents
Keep notes of all cell culture work
Best to have lab coat only for tissue culture work!
Clean the lab often
Disinfect 1-2X/month
Contamination control mats
Recommendations: Working with cells
Don't use antibiotics in culture media
Antibiotics can mask the problem
Use only cell lines confirmed by DNA fingerprinting
Recommendations: BSC
Avoid clutter, use only items you need
Work a few inches inside the front grill, do not block vents!
Change pipet-aids filters regularly, immediately when wet
Keep beaker with disinfectant inside for liquid waste
Clean inside wall often
Recommendations: CO2 incubator
Change to fresh sterile distilled water every 2 weeks
Check for air vents blowing onto the incubator

Notes from Talks Page 1

 Check for air vents blowing onto the incubator
Air ducts can harbor many fungi
Clean the incubator once or twice a month
At least wipe down with 70% alcohol and air dry
Don't forget door and handle
CO2 incubator
Contamination comes from outside- YOU, air
Antimicrobial copper incubators
HEPA filtration
UV and H2O2 are poor choices for incubator decontamination
UV in cell culture incubator - not effective
Do not use H2O2 in the incubator
Cell culture resources
High Throughput Sciences (MIT)
Jaime Cheah jcheah@mit.edu

Notes from Talks Page 2