Fine Needle Aspiration as a Tool To Establish Primary Human Breast Cancer Cultures in Vitro

Abstract

To verify the potential role of fine needle aspiration (FNA) cytology in obtaining malignant cells from primary breast cancer for establishment of a primary breast cancer cell line. In four patients with primary breast cancer subjected to FNA for diagnostic purposes, we attempted to establish primary cultures. We successfully obtained one primary cell line, originating in micropapillary invasive breast carcinoma. FNA material obtained under sterile conditions was centrifuged, and the cell pellet was washed with Dulbecco Modified Medium. The resulting suspension was seeded in 25-cm2 tissue culture flasks. The flasks were maintained with released caps in a 37 degrees C incubator with a humidified atmosphere of 5% CO2 in air. After one week, cells attached to the bottom of the flasks and began proliferating. When a culture became confluent, the cells were treated with 0.05% trypsin/0.02% EDTA in a PBS solution and subcultured. The flasks were observed daily with an inverted microscope, and culture passages were performed weekly. The cell line obtained was named I2FPRW and exhibited morphologic and immunohistochemical features of epithelial cells of mammary origin. The cells were positive for cytokeratins (AE1/AE3 and CK 7), EMA and c-erbB-2. At this writing, this cell line was in the 15th passage of subculturing in the flasks with 10% FBS. In the present study we demonstrated that is possible to establish a breast cancer cell line from material obtained by FNA cytology. FNA seems to be a valuable method of obtaining malignant cells from breast cancer able to grow free of fibroblasts in cell cultures.