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JoVE: Trypsinizing and Subculturing Mammalian Cells (Video Protocol)

Journal of Visualized Experiments

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Trypsinizing and Subculturing Mammalian Cells

Richard Ricardo, Katy Phelan
Molecular Pathology Laboratory, Inc.
0:00 Title
0:37 Introduction
0:56 Trypsinizig and Subculturing Mammalian cells from a Monolayer
4:00 Passaging Cells in Suspension Culture
5:36 Conclusion
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.

Ricardo R, Phelan K (2008). Trypsinizing and Subculturing Mammalian Cells JoVE. 16. http://www.jove.com/index/details.stp?id=755, doi: 10.3791/755
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