Solutions for Successful Cell Culture
Accuracy in cell culture research hinges on creating and maintaining the right environment for your samples. Here are a few tips to ensure that your research yields reliable results.
Preventing Contamination in Your Biological Safety Cabinet
Biological Safety Cabinets (BSCs) serve two main purposes: preventing contamination to protect your samples, and protecting you and your surroundings from your samples. Thermo Scientific BSCs employ fillters and uniform airflows to create an invisible barrier between the outside and inside of the cabinet to protect the user and samples from contaminants such as microorganisms and nucleases.
Go with the flow
When using a BSC, ensure that your samples only encounter contaminant-free air. In addition to using filters, you can minimize interference with the BSC: place everything you need inside the BSC before you start, and avoid fast arm movements as this could cause turbulence which alters the airflow.
Cleaning solutions for BSCs
While bleach is a highly effective germicide, it is corrosive to the steel surfaces of the BSC. Limit your use of bleach to emergencies, such as an infectious spill. For routine disinfection, 70 percent ethanol is effective and safe for steel. Allow the ethanol to air dry for best efficacy.
UV light can kill most microorganisms in about 30 minutes and it can reach areas of the BSC that are difficult to reach, taking some manual labor out of the disinfection process. Make sure to use a BSC with a safety interlock preventing operation of the UV light unless the window is closed.
How Clean is your CO2 Incubator?
Location, location, location
Position your CO2 incubator away from areas of high traffic, but out of neglected corners that may be sources of contamination. Also, make sure it’s sheltered from ventilation and other airstreams that could deliver airborne contaminants.
Cleaning solutions for CO2 incubators
The best way to clean your CO2 incubator is to use mild dish soap in water. Use clear water to rinse and then apply a 2 percent solution of a quaternary ammonium disinfectant, followed by 70 percent ethanol to remove any residues that could harm incubator components or your cells.
The Right Water Purification System for Your Cell Culture Lab
Cell culture labs require ultrapure water that meets or exceeds ASTM Type I standards. Water must be low in ions, organics, bacteria and most importantly endotoxins, which can interfere with cell growth. Ultrapure water systems that incorporate dual wavelength UV light and ultrafiltration technologies are important features to consider when ensuring the best quality of water for successful cell cultures.
While ultrapure Type I water is required for more critical applications in the lab, the lack of ions makes it aggressive and it attacks the stainless-steel components of lab equipment such as CO2 incubators and water baths. ASTM Type II or III water is recommended for these applications. If Type I water is the only source of lab water, it is suggested to add ions and raise the pH to 7-9 with a small amount of a sterile, weak solution of sodium bicarbonate.
Choosing the Right Surface for Your Cell Cultures
Understanding both how surfaces work and the environments they create for your cells is a key component to successful cell culture. When you’re setting up your culture, you’ll need to answer these key questions:
- Will the culture require a hydrophobic or hydrophilic surface?
- Will it benefit from additional coating?
- What harvesting method will you employ?
- Do you want to encourage spheroid formation in your cell cultures?
Your answers will guide you to choose the right cultureware surface. Consider these basics on five surface types when making your selection:
Surface energy treatmentdelivers electronic energy into a cloud of gas, creating energetic oxidation. Depending on which energy treatment is applied — corona or plasma — your cultureware surface will be ideal for different types of cells.
Extracellular Matrix Coating (ECM) uses passive absorption, ideal for cells such as neurons that will not adhere to an energy-treated coating. The ECM coating helps with the binding of tricky cells by replicating the basal lamina substrates on culture surfaces.
Polymer graftuses covalent binding to keep the polymers attached to the polystyrene surface, allowing greater control over the adherence and release of cells.
Untreated polystyreneis a good choice if you need to create a non-adherent environment at a low cost.
Ultra low binding surface coating encourages a spheroid formation by blocking the adsorption of extracellular matrix (ECM) proteins that mediate cell adhesion. With such properties, the cells are unable to attach to the culture dish and instead attach to one another via cell-cell adhesion. As further cells attach, a spheroid is grown.
Choosing the right environment to surround your samples may ensure that you get reliable results in the lab.
Content provided by Thermo Fisher Scientific