If you have ever heard of a HeLa cell, that is an example of an immortalized cell line. In my research, I perform primary culture and also work with cell lines.
Once cells have been cultured, there are three main conditions required for cell growth and survival. Just as humans need food, air, shelter and water to survive, cells can only grow at specific temperature, air composition and nutrient conditions. In the lab, all of the cells that I grow are kept in an incubator at 37°C and 5% CO2. These are standard incubation conditions because they are representative of conditions found in the human body and favorable for the growth of most types of cells.
Media is the liquid in which cells are kept and grown in while in culture. The type of media, and its nutrient composition, changes depending on the type of cell that is used. When I culture the two triple negative breast cancer cell lines, MDA-MB-231 and MDA-MB-iNOS, I use a media known as R.P.M.I 1640. This media was developed to culture human leukemia cells, but is now a standard for many different types of cells. It includes nutrients such as vitamins and amino acids (the building blocks of proteins). For both cell lines, I add 10% fetal bovine serum (F.B.S.) to supply proteins and growth factors for aid in cellular growth. I also add 1% of a combination of two antibiotics, Penicillin-Streptomycin (P/S), to help prevent the growth of unwanted bacteria among my cells.
When working with MDA-MB-iNOS cells, I add a small amount of another antibiotic, puromycin, to the media. i.N.O.S cells exhibit resistance genes to puromycin, but any other microorganisms or cells will not survive in the culture.
