The Importance of Sterility in Cell Culturing: Tips and Tricks
Cell culturing has become a cornerstone in biological research, with its applications ranging from gene editing and protein expression to drug discovery and toxicology studies. The quality of a cell culture critically depends on maintaining sterile conditions throughout the process. To ensure that your cell culturing techniques deliver reliable and reproducible results, it is essential to follow these tips and tricks:
Why is Sterility Critical in Cell Culturing?
The primary goal of cell culturing is to propagate cells in vitro and maintain their biological properties similar to that of in vivo conditions. The growth environment provided in vitro can be influenced by various factors such as the culture medium, supplements, and gases, to mention a few. In addition, cells are prone to microbial or fungal contamination, which can significantly impact the reliability and reproducibility of the results obtained.
Contaminated cultures can lead to misinterpretations, incorrect results, and ultimately, the loss of valuable time and resources. Therefore, sterility is a fundamental requirement for achieving success in cell culture experiments.
Tips and Tricks for Maintaining Sterility in Cell Culturing
1. Cleanliness and Hygiene
Maintaining a clean and hygienic workspace is the foundation for sterile cell culturing. Use alcohol-based disinfectants or UV light to sterilize all instruments, cabinets, and work surfaces before use.
It is also crucial to wear protective clothing such as lab coats, gloves, and face masks that prevent contamination from skin, hair, or saliva.
2. Add Antibiotics and Antifungals
Adding antibiotics and antifungals to your cell culture medium is a common practice to prevent bacterial and fungal contamination. However, it is essential to keep in mind that antibiotics can negatively affect your cells’ health and physiological properties, leading to inaccurate experimental results.
Therefore, it is advisable to use antibiotics only when necessary and in the minimum effective concentration.
3. Maintain Proper Culture Conditions
Consistency in the culture conditions such as temperature, humidity, and CO2 levels is crucial for maintaining the sterility of cell culturing. Changes in the culture environment can create an environment conducive to bacterial and fungal contamination.
4. Avoid Crowding Cells
Overcrowding your culture dish or flask can lead to increased cell waste release, which fosters bacterial and fungal growth. Cell-cell contact within the dish or flask can also lead to cell death and further compromising sterility.
Case Study: Aseptic Techniques in Culturing Stem Cells
The University of Arizona implemented aseptic techniques as the primary means of achieving sterility in culturing adipose-derived stem cells. The study emphasized that stem cells are essential targets for maintaining aseptic techniques due to their low tolerance to microbial infections.
The researchers utilized an ISO-certified cleanroom with laminar airflow to culture the stem cells and disinfected their tools and work surfaces using 70% isopropyl alcohol. The resulting stem cell cultures were healthy and sterile, with no signs of bacterial or fungal contamination.
Conclusion
Maintaining sterility is a fundamental aspect of cell culturing to obtain consistent and reliable results. To maintain sterility, it is essential to keep your workspace clean, wear appropriate protective clothing, use antibiotics only when necessary, maintain proper culture conditions, and avoid overcrowding cells. Adhering to these tips and examples such as the one mentioned in the case study can ensure optimal sterility in your cell culture experiments.
