A rapid and simple method for the purification of tumor cells from ascitic fluid of ovarian carcinoma

Abstract

Rapid isolation of tumor cells growing in clumps from the ascitic fluid of patients with ovarian carcinoma was achieved by harvesting cells from ascitic fluid and subsequently passing them over a 30-microns nylon mesh filter. Single cells and small cell aggregates passed through the mesh, and large cell clumps that had not passed through the filter were isolated by backwashing. Morphologically, the cells in the clumps consisted almost entirely of tumor cells. The clumps were analyzed by immunocytochemistry and only a small proportion of cells (3.1 +/- 0.5% to 8.3 +/- 0.8%) stained with anti-CD45 monoclonal antibody (a panleukocyte marker for cells of hematopoietic origin). Most of the cells in clumps stained positively (90.6 +/- 1.7% to 97.5 +/- 0.5%) with 2G3 (a monoclonal antibody binding to a high-molecular-weight carcinoma-associated antigen). Unfiltered cell populations, however, contained up to 34.4 +/- 2.1% contaminating CD45+ non-tumor cells. This method should be useful for rapidly and easily obtained highly enriched fresh ovarian carcinoma cells from ascitic fluid in a form in which they grow naturally.