Table 1.
Summary of the flow cytometry analyses of selected cell type markers
| Cell model | Passage no | Percentage of positive cells | ||||
|---|---|---|---|---|---|---|
| Vimentin | α-SMA | CK7 | CK18 | CK19 | ||
| MAC-T | 15–22 | 79–93 % | 98–100 % | 0.4–0.9 % | 92–99 % | 0.1–0.2 % |
| bMECUS | 12–15 | 51–87 % | 56–64 % | 35–50 % | 70–88 % | 92–96 % |
| bMECCH | 9–12 | 59–81 % | 69–83 % | 61–62 % | 90–95 % | 75–85 % |
Values show ranges of the percentage of positively stained cells. Data are derived from two to three independent measurements for each cell model. The fluorescence intensity corresponds to the intensity of FITC conjugated polyclonal goat anti-mouse IgG Ab (BioLegend) positively reacting with mouse anti- α-smooth muscle actin (α-SMA) mAb (Novus Biologicals), anti-vimentin mAb (Sigma), anti-cytokeratin (CK) 7 (Dako), anti-CK18 mAb (Sigma), and anti-CK19 (Abcam), respectively. The staining was acquired by counting a minimum of 15,000 events. An IgG1 isotype control staining has been performed to ascertain the reliability of the positive staining. The background fluorescence corresponds to the intensity of FITC conjugated polyclonal goat anti-mouse IgG mAb staining in the presence of the isotype control IgG1 mAb (DakoCytomation,)
MAC-T immortalized bovine mammary epithelial cell line, bMEC US bovine primary mammary epithelial cells isolated from an American Holstein cow at mid-lactation, bMEC CH bovine primary mammary epithelial cells isolated from a Swiss Holstein–Friesian cow at late lactation, Ab antibody