Establishment of pleural effusion cells in culture from a breast cancer patient

Pleural effusion fluid was obtained from a breast cancer patient diagnosed with invasive, poorly differentiated (grade Ill/Ill) ductal carcinoma. The fluid was centrifugea and cells were placed into culture dishes with minimum essential medium, 20% fetal calf serum, 1% glutamine, and antibiotics. After 1 week, cells were proliferating and within 20 days enough cells were obtained to perform immunofluorescence (IMF) studies for intermediate filament (IF) proteins, in order to determine if epithelioid cells were present. Pleural effusion cells in passage one were processed for IMF using a polyclonal antibody to keratin and a monoclonal antibody to vimentin. IMF revealed an elaborate but typical network of keratin IPs. These filaments extended throughout the cell and made contact at the cell periphery with other keratin IF bundles from contiguous cells. Vimentin IFs were present but did not extend to the edges of cells, as did the keratin IFs. Actin filaments were prevalent, arranged in typical bundles and also circumferentially around circular, flat cells. The cultured cells thus appear to be of epithelial origin, representing the metastatic cells. Further tests are necessary to establish cell characteristics and the relationship of nuclear features noted in the original tumor cells (Meek, et al., 1995).