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Durham University

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Publication details for Dr Martin Schröder

M. Schröder & P. Friedl (1997). A protein-free solution as replacement for serum in trypsinization protocols for anchorage-dependent cells. Methods in Cell Science 19(2): 137-147.

Author(s) from Durham


Many animal cell culture media require serum as a component necessary for supporting the growth and survival of the culture. However, serum is of unknown composition, which may vary from lot to lot, and is very protein-rich. Therefore, serum-free media have been developed for the investigation of nutritional demands of animal cells in vitro and the production of therapeutic proteins with animal cells. Despite the fact that there are many published serumfree media for anchorage-dependent animal cells, only little work has been done on the development of serum- and protein-free subcultivation techniques. However, the serum-free subcultivation of animal cells is often a cumbersome attempt resulting in clumping and loss of the major part of the culture. Here we describe a new subcultivation technique for anchorage-dependent cells. Anchorage-dependent cells were first detached from the culture surface by trypsinization, suspended in phosphate buffered saline, 0.02% (w/v) EDTA, 75 mg/l phenol red, and 1% (w/v) pluronic F-68, and centrifuged to separate the cells from trypsin. The use of a trypsin inhibitor was not necessary. Cells were then resuspended in medium and seeded into new tissue culture vessels. No interference of the subcultivation process with the viability of the cells was observed as judged from clonal growth assays.