Fig. 2
EpiProPlus growth media enables large-scale expansion of breast epithelial cells in 2D cultures. a Experimental outline for ex vivo propagation of primary human breast epithelial cells (HBECs) using EpiProPlus media (Lin-, Lineage-depleted; BC, breast cells). b Top panels: representative photomicrographs of HBECs grown in 2D adherent cultures without feeder fibroblasts using EpiProPlus media over five passages. Middle panels: representative FACS plots depicting CD49f and EpCAM expression in ex vivo expanded HBECs. Lower panels: representative photomicrographs of ex vivo expanded HBECs grown in Matrigels as organoids. Total cell numbers (c) and CFC yields (d) per passage for different ex vivo expanded cells (generated from eight different reduction mammoplasty samples) are plotted as individual line graphs. e Representative colony pictures and progenitor cell frequency (luminal and mixed colony forming cell numbers) are plotted for multiple passages. Results are represented as mean ± SEM from at least three biological replicates. Bars represent 400 μm (b) and 1000 μm (e). Representative colony pictures and progenitor cell frequencies were plotted for multiple passages. A, B gated cell populations, CFC colony forming cell, EpCAM epithelial cell adhesion molecule, Lin– lineage depleted, P passage