Section7:Standard HTS Cell Culture Practices to Reduce Contamination Risk

From Assay Guidance Wiki

• When moving things (media bottles, pipet tips, etc.) into a biosafety cabinet (hood) you should wipe them down with 70% EtOH. Be sure to cover the entire object.
• Wipe out hoods before and after use with 70% EtOH. UV weekly. Lysol (3%) weekly.
• Hoods should be completely cleaned at least 3 times a year. This entails taking the surface tray and grills out, washing them and the area beneath them with Lysol then EtOH, and removing any debris found below the tray. Autoclaving the tray and grill are acceptable.
• Bleach vacuum flasks and lines. Change the flask weekly even if not full.
• Wipe out incubators at least once a month with Lysol followed by 70% EtOH. This is a known source of fungal contamination. When opening an incubator, check for fungal growth on the shelves and around the seals.
• Empty biowaste containers regularly, preferably at least twice a week. All waste should be double bagged (bag into container then another bag inside the first).
• Do not carry large stacks of plates or flasks unless you use a cart.
• Wear gloves. Make sure your lab coat is not grimy.
• Bleach any container that has contained cells for a few minutes. The bleached media can be washed down the sink. However, do not open CONTAMINATED containers in the main lab area. If you have a contamination, autoclave it BEFORE opening.
• Fluid delivery lines/ drain lines should be rinsed with 70% EtOH chased with dH2O every day after use. This is a known source of contamination. This would include multidrop heads, multimek lines, MRD8 lines, etc. . If dispensing media with a multidrop, rinse head then autoclave the head. You should keep an autoclaved head in reserve if possible in case of failure of the daily one. If using the Multimek to aspirate or plate cells, rinse the lines and wash station then autoclave the wash station. The autoclaving does not apply to heads used for 384 well delivery.
• If you have a contamination event, DO NOT OPEN IT IN THE MAIN LAB! Contact your supervisor or a cell culture person to help in identifying the contamination and the source. Also, make others using cells aware that you had a contamination event.
• Be careful not to touch pipets, media bottle openings, etc. Touch events are a leading source of contamination in cell culture. If you do happen to touch a pipet discard it and get a new one. If you touch a bottle opening, wipe it immediately with an EtOH swipe and then filter that media through a 0.2um filter apparatus.
• Cleanliness is next to Godliness especially in cell culture. Keep your hood free of unnecessary clutter. Wipe up spills promptly. Try not to sneeze inside your hood. Drips should be promptly cleaned up with EtOH.

Note the sash level limit on your hood. There should be a mark or arrow on one side of the sash (glass) to tell you how high not to go with the sash. Too high will disrupt the air flow and compromise your hoods sterility.