Elimination of the yeastCandida parapsilosis from lymphoid cells and monolayer cells in culture |
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Authors: | Ursula J Behrens Fiorenzo Paronetto |
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Institution: | (1) Immunopathology Laboratory, Veterans Administration Medical Center, 10468 Bronx, New York |
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Abstract: | Summary In our laboratory, airborne yeast contaminants of cell cultures have consistently been of the genusCandida (speciesCandida parapsilosis), which are difficult to control with fungicidal agents. To salvage cell lines that show the presence of this fungus, two
effective methods may be employed. In early stages of infection, the addition of activated mouse peritoneal macrophages (5×105 cells/ml) to the culture medium containing 5 μg Fungizone/ml eliminates all spores by phagocytosis. More heavily contaminated
cultures can be depleted of fungi by density centrifugation on a layer of 38% Percoll. Remaining single spores, often not
detectable by light microscopy, can be removed by the addition of macrophages (2×105/ml) and Fungizone (5 μg/ml) to the culture medium. Contaminated monolayer cells can be freed of blastospores by several washes
with balanced salt solution and subsequent culturing for 4 d in medium containing 10 μg Fungizone/ml without any toxic effects
to the cells. These procedures can rescue valuable cell lines and hybridomas that would otherwise be lost.
This work was supported by Veterans Administration Research Funds. |
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Keywords: | Candida parapsilosis macrophages Percoll |
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