Starting from the thawing of your chosen cell line, we will briefly describe common practices in incubation. When the available space in the cell culture vessel is nearly exhausted (ca. 80 % confluency), the cells need to be transferred to a new container to enable further cell growth. This step also referred to as passaging (or splitting), is necessary to prevent contact inhibition and lack of nutrients do to overpopulation, which may stunt cellular proliferation.

When passaging, adherent cells are detached from their culture vessel - this step is not required for suspension cell lines - and counted to then be seeded in the desired concentration in another container. The generated subculture is then incubated again, performing medium changes every time it is necessary, until the cutoff for confluence is reached once again. Every time splitting is performed, it is possible to isolate excess cells for other purposes, namely cryopreservation, cell-based assays and DNA/RNA/protein extraction. Finally, any remaining cells need to be appropriately disposed of with autoclaves.

Please keep in mind that cell culture conditions differ for each cell type, this page should only serve as an overview of standard cell culture practices and present materials most suited to your work. For detailed protocols always refer to original instruction provided with your specific cell line.