SELECTIVE CELL-CULTURE OF PRIMARY BREAST-CARCINOMA

We have used culture conditions which simulate the microenvironment of breast tumors for the isolation and propagation of primary breast tum or cells in vitro. In this monolayer setup, the mixture of cells disso ciated from primary breast tumors is subjected to self-created gradien ts of oxygen and nutrients as well as metabolic waste and extracellula r pH. The tumor populations isolated under these novel conditions have displayed phenotypic properties characteristic of breast carcinomas, including homogeneous expression of cytokeratin 19, and increased mito chondrial retention of the cationic dye rhodamine 123. Nonmalignant cu ltures from reduction mammoplasty were unable to survive these conditi ons. One tumor population which reached passage 10 was aneuploid for c hromosomes 15 and 17, and displayed a p53 mutation in exon 8. These st udies strongly suggest that the culture conditions described here can suppress the growth of normal breast cells, thereby allowing selective isolation of some populations of slow-growing primary tumor cells in vitro.