Ultrastructure of Breast Cancer Cells in Aggregate Cultures

Abstract

The culture of carcinoma cells in histiotypic aggregates floating free in medium is currently receiving increased attention. The methods used rely on mechanical agitation on rocker or rotary platforms. But mechanical agitation is technically cumbersome and interferes with spontaneous aggregation and organization of the cells by mutual recognition. Epithelial cells were long known of being unable to adhere to agar and we used this phenomenon to grow breast cancer cells in stationary cultures in agar-inlaid, conventional plastic flasks. In this way the cells formed floating aggregates which revealed important features by light and electron microscopy.Corning T25 and T75 flasks were inlaid with 5-6 mm thick layer of 1.5% purified agar boiled in distilled water and sterilized. Once the agar has settled an equal amount of medium (L15 with 10% fetal calf serum but no antibiotics) was added and the flasks incubated for 5 days at 37°C. This allowed time for medium to soak the agar and for contamination to be detected.