Tissue culture studies on pleural effusions from breast carcinoma patients.

SUMMARY The general lack of success that has attended attempts to establish tumor cell lines from solid specimens of breast carcinoma prompted our use of pleural effusions for this purpose. This paper summarizes tissue culture studies on five sequential pleural effusions from each of two patients with breast carcinoma. The morphology of cells in fresh specimens and cultures was documented by light and electron microscopy. Tumor cells were abundant in the fresh fluid specimens and they showed a high degree of viability. Fibroblasts were generally absent. Tumor cells in the fresh fluids and in cultures were morphologically similar to tumor cells cultured from solid breast carcinoma tissue. Nonneoplastic cells were found to survive or proliferate for I to 6 months in culture. Two cell types that grew readily in one of our media (medium BTN) resembled either fibro blasts or mesothelial cells by light microscopy. Electron microscopic studies indicated that the fibroblast-like cells were structurally modified epithelial cells. Our results demonstrate that pleural effusions provide a fertile source of tumor cells from which tumor cell lines can be established. Fibroblasts are rare or absent, although other nonneoplastic cells are usually present and can interfere with attempts to obtain pure cultures of tumor cells. We have now studied over 40 pleural fluid specimens, some of them sequential aspirates from the same patient. The results presented here are representative of our experience. They are mainly based on findingsin 10 specimens, 5 obtainedfrom each of 2 patients at intervals of I to 2 months. The tumor cells observed and 2 of the main nonneoplastic cells (fibroblast like and mesothelial-like) are described in detail. MATERIALS AND METHODS Pleural fluids were obtained aseptically in vacuum bottles by thoracentesis and treated as outlined in Chart I. After trying out numerous media considered adequate in the establishment of other types of tumor lines, we derived our own combination of components believed essential. Two types of media were used with all cultures. In addition, modifications of these 2 media including minor changes in the amounts of serum, hormones, and carbohydrates were also used. Media INTRODUCTION Numerous attempts to culture human breast carcinoma cells have been made over the last 20 years, with limited success (I, 2, 4, 7, 10). A primary cause of failure has been the presence of fibroblasts that occur in practically all breast tissue specimens. These ubiquitous cells proliferate rapidly and tend to smother the …