Cell culture is a critical function of many life science laboratories, but how often do we stop to think about the best practices that could make our job a bit easier or our time at the bench more productive.

In this Best Practices article, we spoke with Dr. Kyung-A Song, NA Scientific Support Manager, Corning Life Sciences about the steps we can take to improve our time at the bench. We discussed the importance of ergonomics, bench set up, cell culture set up, time saving opportunities, and having the right tools and products to support these efforts.

Clean and Organized Workspace 

This speaks for itself, but sometimes we all need a reminder about the importance of a clean and organized bench. We all know that sterility is so important, so having a disorderly bench means that not only could you contaminate something unintentionally, but you could also contaminate your culture.

One area that can be overlooked is the incubator. It must be very clean, and this means it should have a regular maintenance schedule and the water tray used for humidity should be regularly and thoroughly cleaned as it is a common source of contamination.

Supplies at the Ready

Going along with a clean and organized bench, having the necessary products and tools, including plates, dishes, and flasks, cell culture media, and liquid handling tools, such as a pipets and pipet controller, helps to make you more efficient so you don’t need to leave the bench during your work. Additionally, you only want to have the consumables and devices necessary on the bench so you don’t have any additional material that could be contaminated or could introduce contamination.

Bench Layout and Ergonomics are Key

It is important to think about how you will lay out your workspace. Simple things like if you are right- or left-handed need to be considered so you can set up your space to accommodate your needs. This in turn will make your work more efficient and can prevent spills or other problems that may happen if you are routinely reaching over your bench to grab something you need on the other side.

To support this, Corning offers convenient liquid handling instruments, streamlining daily laboratory tasks, and mitigating the risk of contamination. The portable Stripettor® Ultra™ pipet controller accompanied by an ergonomic stand, facilitates easy placement with or without the serological pipet. Even if there are residual liquid remnants in the pipet, the stand ensures a slight tilt, preventing the liquid from infiltrating the device and causing contamination.

For dispensing micro-volumes, it is recommended to use Single- or Multichannel pipettors in conjunction with filtered pipet tips. The extensive range of pipettor stands (linear and carousel) simplifies the storage of pipettors, ensuring their accessibility wherever needed. Proper storage of pipet tips in closed-lid racks is crucial to minimize the risk of contamination. You can also utilize Corning® Lambda™ EliteTouch™ Pipettors and DeckWorks pipet tips in racks so they can be easily accessible. This comprehensive approach by Corning aims to enhance efficiency and maintain a pristine laboratory environment for researchers.

Another way to make your space more work friendly is by using pre-racked conical tubes, which reduces handling of the tubes and makes them easier to use.

Setting Up Your Cell Culture

When setting up your cell culture first and foremost, you need good cell material. It is critical that you start your culture devoid of any contaminants of biological origins such as viruses, mycoplasma, etc.

From there, most researchers follow a standard protocol. For example, in adherent cells with plates, the process would be:

1. Take out vial from vapor phase nitrogen tank.
2. Thaw the vial in a 37-degree water bath.
3. Remove all the freezing media by centrifugation because it contains DMSO and that has cytotoxic properties. Then add a washing buffer and spin it down again to a pellet so you can remove all the freezing media completely.
4. Suspend your cell pellet in the completed culture media that you are going to use for cell growth and selection. Corning has a wide selection of media to use depending on the cell type and specific research goals.
5. Seed your cells and once the cells attach onto the culture flask for 3-4 days later, you aspirate the old media and give the cells fresh media.
6. Check the cell viability and wait for them to grow up to 80-85% of cell confluency.
7. For harvesting the cells, aspirate the old media and add washing buffer and aspirate it again. Add dissociation reagent like Trypsin and incubate the cell at 37°Cin the incubator supplied with 5% of CO₂ for a short amount of time.
8. After the incubation, cells need to inactivate the disassociation reagent with serum containing media and give the cells another washing step.
9. If you want to do another passage, you can split your culture or you can prepare it for freezing by suspending the cells in freezing media and placing them back in cryogenic storage.

Larger Scale Projects

For larger scale projects, such as preliminary steps for manufacturing you can scale up in vessels, such as Corning’s HYPERFlask® or HYPERStack® cell culture vessels  These are very space efficient because you can remove the airspace above the media entirely because the cells can grow in those on oxygen permeable or gas permeable membranes that allow for a very densely packed or compact cell growing system.

For example, the HYPERFlask vessel has a footprint of a T175 flask but has 10 layers to it, so you can grow 10 times as many cells (1,720 cm² growth area) in the same space as you would with one T175. This also saves time and space by reducing the processing time and incubator storage space by handling one flask as compared to 10 traditional 175 cm² flasks. The HYPERFlask vessel is also automation compatible and is bar coded and compatible with the The Automation Partnership SelecT™ and CompacT™ SelecT™ cell culture systems thereby enabling even more efficiency.

You can scale up culture in the HYPERFlask for the HYPERStack where you have 36 layers, so you have 5X the growth surface area of a traditional cell culture vessel of comparable footprint. It is a closed system, so you have a significant reduction in the risk of contamination with no open fluid manipulations. It is scalable with multiple size offerings to support scale up and scale out. It offers an ergonomic design for easier manipulation and handling.

Time Savers

There are lots of ways to make culture more efficient and things that can help conserve time and effort.

Larger Volume Liquid Handling Options

Larger volume liquid handling is important for stacked vessels because you need to transfer larger amounts of media. The Corning Stripette serological pipets for liquid transfers between 5 to 100 milliliters, help with this because they can transfer a larger volume of media and it takes less time and is less tiring for the operator. They come in a wide range of sizes and different packaging options, bulk packaging or individual for sterility. Corning also offers a wide range of pipette tips that come in different configurations such as filter or non-filtered and size ranges from 5 microliters to 1000 microliters.

Specialized Microplates

Corning offers a variety of microplates that offer built-in time-saving features.

• Precoated microplates with different extracellular matrices, so you don’t need to spend time coating the plates.
• Spheroid microplates specifically designed for generating and analyzing 3D multicellular spheroids in the same microplate. The Ultra-Low Attachment (ULA) surface enables uniform and reproducible 3D multicellular spheroid formation. The black opaque microplate body shields each optically clear, round bottom well from well-to-well crosstalk. This allows you to culture and assay spheroids in the same microplate without the need for transfer to a new microplate.
• High content screening microplates are designed for high optical quality and are ideal for performing high-content cell-based assays using imaging systems. They provide a flat and optically clear surface that reduces autofocus time, increases throughput, and is ideal for cell growth.

Automated Cell Counting

Automated cell counting is much faster and less user-dependent than manual cell counting, but the cost of disposable counting slides has led many labs to continue with manual counting.

The  Corning Cell Counter is the first automated cell counter that combines the best of both. It is fast due to its online image processing, accurate because of its cloud-based machine learning algorithm and low cost because it works with common reusable glass hemocytometer, so no consumables are required.

Sustainable Products

Along with all we have discussed about setting up the ideal lab, we should also discuss the importance of sustainable products. Corning has launched a new EcoChoice™ program with products that are produced, packaged and/or distributed in an environmentally friendly manner. These products follow the United States FTC Guidelines and means that all product sustainability statements are specific, evidence-based, and traceable.

Corning EcoChoice Program Products must meet one or more of the following criteria:

• Recycled content (pre-consumer or post-consumer)
• Source reduction
• Intensification: products designed to enable greater cell production in a smaller footprint, resulting in less plastic per unit of output
• Renewable energy/RECs
• Certifications

For example, Corning has redesigned their 75cm² Corning Cell Culture Flask to reduce product plastic by 23%. The new rounded shoulders of the flask also allow for easier laboratory usage, making it a win for scientists and the environment.

About the Expert

Dr. Kyung-A Song, NA Scientific Support Manager, Corning Life Sciences

Dr. Song graduated from Sungkyunkwan University with a Ph.D. in molecular biology and cancer immunology. She completed her postdoctoral fellowship in pharmaceutical research and drug discovery at Virginia Commonwealth University. She has extensive experience in a variety of cell culture techniques including isolation and culture of primary patient cells, has worked with xenograft models and many cancer cell lines, and has expertise in in vitro cell based functional assays and in vivo experiments. She is currently the NA Scientific Support manager for Corning Life Sciences, where she provides trainings and webinars for technical information and its applications in the Corning Life Sciences extensive portfolio.