Improved medium and culture conditions for clonal growth with minimal serum protein and for enhanced serum-free survival of Swiss 3T3 cells |
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Authors: | Gary D Shipley Richard G Ham |
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Institution: | (1) Department of Molecular, Cellular and Developmental Biology, University of Colorado, 80309 Boulder, Colorado |
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Abstract: | Summary Improved culture conditions have been developed that will support clonal growth of Swiss mouse embryo 3T3 cells at concentrations
of serum protein as low as 125μg/ml. Survival of the cells under completely protein-free conditions also is enhanced greatly.
The improvements that made these results possible include: (a) use of medium MCDB 402, which was developed specifically for
Swiss 3T3 cells by adjusting the concentrations of all components of Dulbecco's modified Eagle's medium to optimum values
for clonal growth with minimal serum protein and by adding other nutrients such as trace elements and “nonessential” amino
acids that were not in the original formula; (b) use of culture surfaces that are coated with a positively charged polymer,
poly-d-lysine; and (c) use of gentle low temperature trypsinization technique that minimizes cellular damage and the need to neutralize
residual trypsin.
Portions of this work were reported at the Thirtieth Annual Meeting of the Tissue Culture Association in Seattle, Washington.
This work was supported by Grant CA-15305 from the National Cancer Institute |
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Keywords: | Swiss 3T3 cells survival clonal growth MCDB 402 serum protein |
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