Primary cell culture is an expanding technology for several biomedical applications like tissue culture, 3D Cell Culture, bioscaffolds, 3D bioprinting, organoid culture etc. Researchers have been utilizing this technology as an efficient ethical alternative to animal experiments and as a physiologically in vivo-mimicking alternative to culturing continuous cell lines. We, at Kosheeka, believe that cell culture practice and maintenance can enhance the applications and efficacy of primary cell culture manifold and therefore, here are some simple tips for Primary Cell Culture maintenance that will hopefully help the budding researchers.
Maintain Your Hood
The biosafety cabinet should be turned on for at least 15 minutes before the work in the hood starts. This is to ensure proper clean air flow. The air flow vent opening should be left uncovered to ensure proper flow. Moreover, before work starts, a 10-15 minutes of switching on the UV light ensures that no contamination in the hood stays. UV light is harmful for your eyes and skin and therefore, keep it off while using the hood. After switching off the UV, always wipe the hood surface clean with 70% ethanol or IPA before starting work.
Keep Your Contaminants Away
Like we suggested in the previous point, wipe down your working surface of the hood with 70% IPA or ethanol and also wipe everything that gets inside the hood, including your glove-wearing hands. Remove rings and other ornaments on your body before working in a cell culture lab. Also avoid talking unnecessarily or touching your skin or hair while inside the culture room. All these may sound cliché and trivial but every small step helps in reducing the chance of bringing contaminants from outside in the form of aerosols or microbial contaminants. Avoid moving things in and out of the hood while working on your cell culture and always plan your experiments ahead.
Keep The Hood Tidy And Clean
Hoods are not for storage! Cluttering your hood for work increases contamination risks and makes it more difficult for air flow and working ease by reducing working area and making it difficult for you to clean the hood surface. Just take what you need inside the hood area by planning it beforehand and keep your tubes and bottles closed when not in use, while working. If media spills or any spills occur inside the hood, they should be immediately wiped clean with 70% ethanol or IPA with sterile tissue papers and discarded in the small beaker (used as bin inside the hood for pipette tips). Moreover, instead of using the whole bottle of any solution or media, aliquot the necessary amounts in falcon tubes for ease and keep the main bottle closed to avoid any chance of contamination of the stock solution.
Care For Your Cells In The Hood
Do not let your cell culture flasks stay for too long outside the incubator. pH maintenance is a big issue without incubator and growth is affected due to disturbance of the preferred temperature conditions. Primary cell cultures should be handles with utmost care and without a sterile incubator, conditions might be harmful for the cell viability. Keep the cells inside the incubator while you are preparing other media and solutions so that you can avoid clutter and also help your cells to be happy and healthy. Also ensure that unless specified, temperatures of washing buffer like PBS should be similar to the temperature of the cells to avoid any mechanical harm towards the cells.
Check Cell Morphology And Numbers
A light microscope should be available close to the hood to have the least distance between working on your cells and viewing them. One should also know the proper morphology of the cells that they are working with for ensuring proper maintenance of the cells. This helps in detecting cross-contamination and microbial contamination.
When starting a new primary cell culture, a good practice in maintaining cell numbers is to grow the cells, aliquot them in a number of cryovials, cryopreserve them in liquid nitrogen and use just one of those vials to propagate passages. This ensures that if any issues arise in your cell culture passage, you can always lean back on the original stock of cells.
Hopefully, these simple lab tips for primary cell culture will help you enhance your culture practice in the lab settings and if you are ready to start your primary cell culture, Kosheeka is here to make it easy for you! For procuring species-specific and tissue-specific primary cells, contact us at firstname.lastname@example.org