Behrens U J, Paronetto F
In Vitro. 1984 May;20(5):391-5. doi: 10.1007/BF02619584.
In our laboratory, airborne yeast contaminants of cell cultures have consistently been of the genus Candida (species Candida 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 X 10(5) cells/ml) to the culture medium containing 5 micrograms 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 X 10(5)/ml) and Fungizone (5 micrograms/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 micrograms Fungizone /ml without any toxic effects to the cells. These procedures can rescue valuable cell lines and hybridomas that would otherwise be lost.
在我们实验室中，细胞培养物的空气传播酵母污染物一直是念珠菌属（近平滑念珠菌），用杀真菌剂很难控制。为挽救显示存在这种真菌的细胞系，可采用两种有效方法。在感染早期，向含有5微克/毫升两性霉素B的培养基中添加活化的小鼠腹腔巨噬细胞（5×10⁵个细胞/毫升），通过吞噬作用消除所有孢子。污染更严重的培养物可通过在一层38%的 Percoll上进行密度离心来去除真菌。剩余的单个孢子通常在光学显微镜下无法检测到，可通过向培养基中添加巨噬细胞（2×10⁵/毫升）和两性霉素B（5微克/毫升）来去除。污染的单层细胞可通过用平衡盐溶液洗涤几次，然后在含有10微克/毫升两性霉素B的培养基中培养4天来去除芽生孢子，且对细胞无任何毒性作用。这些程序可以挽救否则会丢失的有价值的细胞系和杂交瘤。
