ANNEX 6: Cell Culture Room
Contact Person: Simon Henriet
The Sars Centre's guidelines are based on the National Institutes of Health's publication "Biosafety in Microbiological and Biomedical Laboratories", known as BMBL. For more information please reference http://www.cdc.gov/biosafety/publications/bmbl5/index.htm
Brief guidelines for working with cell cultures
Only authorised personnel are allowed to use the room. Group Leaders are responsible for ensuring the training and authorisation of lab members. For detailed instructions regarding safety, rules and best practices in the cell culture room, please refer to the facility handbook.
- Only cell cultures which have been approved by the contact person may be cultivated, and under no circumstances must bacteria be brought into the cell lab.
- Wear gloves when working with potentially infectious materials, contaminated surfaces or equipment. Two pairs of gloves may be appropriate.
- Use face protection against anticipated splashes or sprays of infectious or other hazardous materials. If possible, do such work only inside of hoods.
- A lab coat must be worn at all times.
- Disposable shoe covers or clogs (only used in cell lab) must be always be used in the cell culture room. Remember to remove these upon leaving the cell lab.
- Wash your hands after handling viable materials, after removing gloves, and before leaving the laboratory.
- Contaminated cell cultures must be poured into a new glass bottle and autoclaved immediately! Pour a little soap into the cell bottle, replace the cap and autoclave.
- Working with approved viruses requires separate work procedures. Consult the contact person.
- Work surfaces must be decontaminated on completion of work, and after any spill or splash of viable material.
- All cultures, stocks, and other regulated wastes must be decontaminated before disposal by autoclaving. Transport waste to the autoclave in a durable, leak proof container.
- Disposable sharps must be carefully placed in puncture-resistant containers used for sharps disposal. Non-disposable sharps must be placed in a hard-walled container for decontamination by autoclaving.
- When working in the LAF bench, all leftover media/cells/liquids must be poured into a container. This must be autoclaved when full (at least every two weeks).
- Throw away all disposable materials (such as pipette tips, plastic Pasteur pipettes, etc) in the metal bins for autoclaving.
- Gloves and paper towels etc. can be disposed of in the yellow waste boxes.
Brief guidelines for working with retroviruses
Viruses can be used for gene delivery into mammalian cell culture. The viruses currently used at the Sars Centre are not considered pathogenic (they can potentially infect human cells and integrate their DNA, but they cannot proliferate).
- Manipulation of viruses must be done exclusively in the laminar flow hood; the shield will prevent accidental spray of virus solution into your eyes; the laminar air flow helps to keep the surface dry and reduce the risk of virus survival in aerosol.
- Use Barrycidal (a liquid germicide) spray to clean the bench before and after working with the virus suspension, then wipe with ethanol.
- Use Barrycidal spray to clean any spill from virus-containing liquid.
- A special bin filled with Barrycidal is present in the laminar flow hood for disposal of contaminated tips and eppendorfs.
- As these retroviruses are poorly resistant in aerosol, infected cells can be grown in the CO2 incubator.