As cells reach confluency, they must be subcultured or passaged. This video will demonstrate a procedure for subculturing both adherent and suspension cells.
Date Published: 6/12/2008, Issue 16; doi: 10.3791/755
Keywords: Basic Protocols, Issue 16, Current Protocols Wiley, Cell Culture, Cell Passaging, Trypsinizing Cells, Adherent Cells, Suspension Cells
Ricardo, R., Phelan, K. Trypsinizing and Subculturing Mammalian Cells. J. Vis. Exp. (16), e755, doi:10.3791/755 (2008).
As cells reach confluency, they must be subcultured or passaged. Failure to subculture confluent
cells results in reduced mitotic index and eventually in cell death. The first step in subculturing is to detach cells from the surface of the primary culture vessel by trypsinization or mechanical means. The resultant cell suspension is then subdivided, or reseeded, into fresh cultures. Secondary cultures are checked for growth and fed periodically, and may be subsequently subcultured to produce tertiary cultures. The time between passaging
of cells varies with the cell line
and depends on the growth rate.
The complete text protocol for this experimental approach is available in Current Protocols in Cell Biology.
Subscription Required. Please recommend JoVE to your librarian.
The authors have nothing to disclose.