Figure 2.

High enrichment of mammary epithelial cell for primary culture in two-dimensions. (A) Brightfield images of mammary epithelial cells obtained after purification compared with the cells eliminated during the purification process. Scale bars = 300 μm. (B) Western blot analysis of purified cells normally discarded with differential plating (DP) compared to purified epithelial cells (Pur). Protein expression of the cytokeratin 8 and 18 (CK8/18), a marker of epithelial cells, and vimentin, a marker of fibroblasts, after three and six days in two-dimensional culture. S6 was used as the loading control. (C) Immunofluorescence showing the expression of CK8/18 (green) and vimentin (red) at three or six days in two-dimensional culture. Nuclei were stained with DAPI (blue). Scale bars = 300 μm and 75 μm. (D) Ratios of positive cells for CK8/18 or vimentin per the total number of cells (counts of nuclei) in percentage. Data are shown as mean ± SEM of one representative experiment (n = 6 images per condition). **p < 0.01; ***p < 0.001.