Culture-adapted CTC Isolation and Maintenance

Culture-adapted primary CTCs were obtained from peripheral blood from women with luminal B metastatic breast cancer. CTC isolation was performed using the CTC-iChip.[1 (link), 16 (link), 23 (link)] Culture-adapted CTCs were maintained in 6-well ultra-low adhesion plates (Corning CLS3471). Culture media and supplements were obtained from Life Technologies and consisted of RPMI 1640 medium (485 mL), EGF (10 μg), bFGF (10 μg), B27 (10 mL) and 100x Antibiotic-Antimycotic (5 mL). CTCs were maintained in a 5% CO₂/5% O₂/90% N₂ incubator and subculture was performed after cells reached a density of ~500k cells/well. Cells were subcultured at a 1:3 ratio.