Cell culture is the growth of cells from an animal in an artificial, controlled environment. Cells are removed either from the organism directly and disaggregated before cultivation or from a cell line or cell strain that has previously been established.
Certain culture conditions depend on the cell type; however, each culture must consist of a suitable vessel with a substrate or medium that supplies the nutrients (such as amino acids, carbohydrates, vitamins, and minerals), growth factors, or essential hormones for culturing cells. Gases (O2, CO2), and physicochemical environment (pH, osmotic pressure, temperature) also play an important role to regulate the proper cell growth in an artificial environment
Cell culture is an amazing tool that allows for easy control and manipulation of all physiochemical and physiological cell factors, such as temperature, osmotic pressure, pH, gas, hormones, media, and its supplements. Media supplements help to optimize cell growth for specific applications depending on the chosen tissue or cell type. To be successful in cell culture, it is essential to remain in a contamination-free environment (bacteria, fungi, mycoplasma …. etc.), so following aseptic techniques ensure that no microorganisms enter the cell culture.
In this protocol, adherent cell lines will be first thawed from liquid nitrogen and seeded in a cell culture flask. Following 24 hours, cells will be visualized by light microscope for confluency, in order to define the best time for sub-culturing and scaling-up of cells. Once the cells exceed 90% confluency, they will be sub-cultured and passaged into other culture vessels. For long-term storage, cells will be frozen in a special freezing medium before storing them in liquid nitrogen.
