We are going to set up a new cell culture room. What are the necessary procedures to avoid contamination?
If you already have Biosafety Cabinets (hoods), the next step is to get them Certified for operation. Ensure that you have the correct positive ventilation in the room itself based on the area size. Sanitize all surfaces in the room and particularly within the hoods themselves. Once this is done try doing inconsequential work in the hoods, incubate and examine for contamination. Once operational, always use standard GLP and aseptic techniques.
If you already have Biosafety Cabinets (hoods), the next step is to get them Certified for operation. Ensure that you have the correct positive ventilation in the room itself based on the area size. Sanitize all surfaces in the room and particularly within the hoods themselves. Once this is done try doing inconsequential work in the hoods, incubate and examine for contamination. Once operational, always use standard GLP and aseptic techniques.
Yes, BSC will help a lot, get certification to ensure the filter works fine. Also, BSC has many types, depending on what type of BSL you working with. Most of medical research labs will need to have the type II BSC.
When you think that you are ready to start the real work with the cell cultures try to test your hoods and CO2 incubators for reliability. Prepare enriched culture mediums, usually with 10-15% serum, pour into Petri dishes, test them for possible contamination under the microscope and then leave them opened in different parts of the hood or incubator for 1-2 h. Switch on the air flow. Incubate the plates for 24-48 h and test under the microscope for contamination. If do not notice anything suspicious you can start your cell cultures.
but make sure we test our culture with mycoplasma kit first before conducting the test to ensure our stock free from contaminant..
the answers given above are all great. from mine experience the hood for cell culture experiment should be positioned well enough to prevent excessive movement of people across the hood. More importantly, ensure that the hood is always cleaned before and after use esp. after drips of serum and media.This could be done by wiping the surface of hood with 75% ethanol and UV for at least 30 min. Also, the water in the incubator should be distilled and container cleaned periodically with 75% ethanol. The same goes to the water bath used in warming media or thawing cells. This helps to prevent Legionella infection.
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