Cell dissociation is the process during cell passaging where cells are detached from the treated surface to create suspensions. These suspensions are important for reseeding for subculture, cell counting for analysis, and cell propagation. There are various proteolytic enzymes that are used to detach cells from the adherent substrate, including trypsin.
Trypsin is a member of serine protease family frequently used to detach cells. The optimum activity for trypsin occurs at 37 °C, so pre-warmed trypsin is often used to speed up cell detachment. However, long-term incubation with high concentrations of trypsin can damage cells by striping surface proteins and killing the cells.
Trypsin is tolerated by many cell types. However, proteomic studies and experiments requiring serum-free cultures often avoid using trypsin. Based on the application and cell type, various constituents and concentrations of trypsin can be employed. Some of the attributes are discussed below.
This protocol is performed while maintaining aseptic environment, such as a laminar bio monitored hood.
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