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Biology

Trypsinizing and Subculturing Mammalian Cells

Published: June 12, 2008 doi: 10.3791/755

Summary

As cells reach confluency, they must be subcultured or passaged. This video will demonstrate a procedure for subculturing both adherent and suspension cells.

Abstract

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.

Protocol

The complete text protocol for this experimental approach is available in Current Protocols in Cell Biology.

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Disclosures

The authors have nothing to disclose.

Tags

Trypsinizing Subculturing Mammalian Cells Confluency Passaging Mitotic Index Cell Death Detachment Primary Culture Vessel Trypsinization Mechanical Means Cell Suspension Reseeding Fresh Cultures Secondary Cultures Growth Feeding Tertiary Cultures Time Interval Cell Line Growth Rate
Trypsinizing and Subculturing Mammalian Cells
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Cite this Article

Ricardo, R., Phelan, K. Trypsinizing More

Ricardo, R., Phelan, K. Trypsinizing and Subculturing Mammalian Cells. J. Vis. Exp. (16), e755, doi:10.3791/755 (2008).

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