Previous ultrastructural and biochemical studies on ductal infiltrating carcinomas (d.i.c.) of human breast have suggested that interactions between the tumour cells and extracellular collagenous matrix are important for the invasive behaviour of these neoplastic cells (Minafra et al., 1984a(Minafra et al., ,b, 1988Pucci Minafra et al., 1985. To examine this further we have attempted to establish a breast carcinoma cell line in vitro which retains many of the properties observed in vivo. To date the majority of breast cancerderived cell lines have been obtained from secondary tumours and pleural effusions (Dobrynin, 1963;Soule et al., 1973;Trempe & Fogh, 1973;Engel & Yung, 1978;Monaghan et al., 1985). Although a cell line derived from a primary breast carcinoma has recently been reported (Vandewalle et al., 1987) the relative paucity of cell lines derived from primary carcinomas (Hackett et al., 1977;Nordquist et al., 1975; Lasfargues et al., 1978;Rudland et al., 1985) may be related to the technical difficulties associated with the extraction of viable tumour cells from surrounding stroma. Here we report the isolation, establishment and characterisation of a continuous line of neoplastic cells isolated from a primary duct infiltrating carcinoma of human breast (8701-BC). These cells have been grown in monolayer cultures for more than 100 passages and have retained morphological characteristics similar to those of the original tumour. Materials and methods Tissue fractionationTumour fragments obtained from surgical operations and histologically diagnosed as ductal infiltrating carcinoma were washed under sterile conditions with a Ca2+ and Mg2+ -free balanced salt solution (BSS-CMF). Specimens to be used for cell culture were taken from the core of the tumour, cut into very small pieces and pre-incubated in BSS-CMF with gentle stirring at 37°C for 15 min. 8701-BC cell line was derived from a G2-G3 duct infiltrating carcinoma obtained from a 72-year-old patient, with extensive lymphonodal infiltration. Histological grading was evaluated according to Bloom & Richardson (1957).Cell culture Cells of primary cultures were grown in minimum essential medium (MEM) with Earle's salts (Biochrom) supplemented with 20% (v/v) FCS (Difco) and 10% (v/v) TPB (Difco). A cell density of 1-2 x 10s cells ml-1 was used to seed primary cultures in 5ml flasks. At confluency cells were subcultured following detachment by exposure to trypsin 0.5% (w/v) and Versene 0.04% (w/v)
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