Fine needle aspiration as a tool to establish primary human breast cancer cultures in vitro

OBJECTIVE: To verify the potential role of fine needle aspiration (FNA) cyt ology in obtaining malignant cells from primary breast cancer for establish ment of a primary breast cancer cell line. STUDY DESIGN: In four patients with primary breast cancer subjected to FNA for diagnostic purposes, we attempted to establish primary cultures. We suc cessfully obtained one primary cell line, originating in micropapillary inv asive 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-cm(2) 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 co nfluent, the cells were treated with 0.05% trypsin/0.02% EDTA in a PBS solu tion and subcultured. The flasks were observed daily with an inverted micro scope, and culture passages were performed weekly. RESULTS: 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. CONCLUSION: In the present study we demonstrated that is possible to establ ish a breast cancer cell line from material obtained by FNA cytology. FNA s eems to be a valuable method of obtaining malignant cells from breast cance r able to grow free of fibroblasts in cell cultures.