Last updated on Aug 27, 2024

What are the best practices for maintaining a sterile cell culture?

Cell culture is a widely used technique in bioengineering, where cells are grown and manipulated in controlled conditions. However, cell culture can be easily contaminated by microorganisms, such as bacteria, fungi, and viruses, that can affect the quality and reliability of the results. Therefore, maintaining a sterile cell culture is essential for any bioengineering project. In this article, you will learn some of the best practices for keeping your cell culture free from contamination and ensuring its optimal performance.

1 Choose the right materials

One of the first steps for maintaining a sterile cell culture is to choose the right materials for your experiment. You should use sterile plastic or glassware, such as flasks, pipettes, and tubes, that are designed for cell culture and have tight seals. You should also use sterile media, buffers, and solutions, and avoid reusing or sharing them with other experiments. You should check the expiration dates and storage conditions of your materials, and discard any that are cloudy, discolored, or have signs of microbial growth.

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Dan Storey

Chief Executive Officer at CMDC Labs
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Select high-quality, sterile materials such as culture media, reagents, and labware to minimize contamination risks. Ensure that all materials are stored and handled properly, using single-use, pre-sterilized items whenever possible. Using the right materials is the first step in maintaining a sterile environment for your cell culture.

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2 Work in a clean environment

Another important step for maintaining a sterile cell culture is to work in a clean and organized environment. You should use a biosafety cabinet or a laminar flow hood, which are enclosed spaces that provide filtered air and prevent contamination from the surrounding air. You should also clean and disinfect your work area, equipment, and hands before and after each experiment, using appropriate agents, such as ethanol, bleach, or autoclave. You should also wear protective clothing, such as gloves, masks, and lab coats, and avoid touching your face, hair, or clothes while working.

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William W.

MD/PhD Candidate | Harrington Scholar
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The best pieces of advice I have been given are to assume every external surface is contaminated and if you are not 100% sure that your pipette tips are sterile, toss it and grab a new one. I would rather lose $0.25 on a serological pipette than 4 hours of work and $300 of reagents because I wasn't sure!

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Dan Storey

Chief Executive Officer at CMDC Labs
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Conduct all cell culture work within a certified biosafety cabinet or laminar flow hood. Regularly clean and disinfect the workspace before and after use, and ensure that the airflow system is functioning correctly. A clean environment significantly reduces the risk of introducing contaminants into your cultures.

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3 Follow aseptic techniques

Aseptic techniques are essential methods to prevent the introduction of microorganisms into your cell culture. Whenever you open, close, transfer, or manipulate your cell culture, you should follow these techniques. For instance, flame the necks of your flasks or bottles before and after opening them with a Bunsen burner or an alcohol lamp to kill any microorganisms. Additionally, use sterile pipette tips and change them after each use to avoid cross-contamination. It is also important to minimize the exposure of your cell culture to the air by keeping lids or caps closed and working quickly and efficiently. Lastly, label your cell culture clearly and keep a record of its origin, passage number, and any treatments or modifications to avoid confusion or mix-ups.

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Dan Storey

Chief Executive Officer at CMDC Labs
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Consistently practice aseptic techniques, including proper handwashing, wearing gloves and lab coats, and using sterile instruments. Avoid unnecessary opening of containers and minimize movements that could disturb the airflow. Strict adherence to aseptic methods is crucial for preventing contamination during cell culture procedures.

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4 Monitor your cell culture regularly

The final step for maintaining a sterile cell culture is to monitor it regularly for any signs of contamination or changes. You should observe your cell culture under a microscope, looking for any abnormalities in the morphology, density, or behavior of your cells. You should also check the color and pH of your media, looking for any turbidity, precipitation, or acidity that may indicate microbial growth. You should also perform quality control tests, such as staining, PCR, or ELISA, to confirm the identity and purity of your cell culture. If you detect any contamination or problems, you should discard your cell culture and start a new one, using fresh materials and following the best practices.

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Dan Storey

Chief Executive Officer at CMDC Labs
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Inspect cultures frequently for signs of contamination, such as unexpected changes in pH, turbidity, or the presence of foreign particles. Regularly check the sterility of media and reagents, and if contamination is detected, promptly discard the affected cultures and materials. Regular monitoring ensures early detection and prevention of contamination spread.

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5 Here’s what else to consider

This is a space to share examples, stories, or insights that don’t fit into any of the previous sections. What else would you like to add?

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Dan Storey

Chief Executive Officer at CMDC Labs
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Maintain meticulous records of all cell culture procedures, including the source of materials and any deviations from standard protocols. Regularly train and retrain personnel in sterile techniques and the use of equipment. Implementing a strong culture of sterility awareness and documentation helps to maintain long-term sterility and reproducibility in cell culture work.

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What are the best practices for maintaining a sterile cell culture?

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1 Choose the right materials

2 Work in a clean environment

3 Follow aseptic techniques

4 Monitor your cell culture regularly

5 Here’s what else to consider

Bioengineering

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