Characterization of Primary Breast Cancer Cells
| Patient No. . | Age (yr) . | Source . | Immunocytochemistry Results . | Western Blot . | Termination of Culture . | |||
|---|---|---|---|---|---|---|---|---|
| Anticytokeratin (AE1/3+ KL-1) . | Anticytokeratin (MNF116) . | Antiepithelial (Ber-Ep4) . | Anticalretinin (08-1211) . | erbB2 Protein (21N) . | ||||
| 1 | 68 | Ascites | + | + | + | − | High | d 80 |
| 2 | 61 | Ascites | + | ND | + | ND | Int | d 90 |
| 3 | 48 | Pleura | + | + | + | − | High* | d 60 |
| 4 | 84 | Pleura | + | + | + | − | Int | d 80 |
| 5 | 49 | Ascites | + | + | + | − | High | d 80 |
| 6 | 73 | Pleura | + | + | + | − | No/low | d 130 |
| Patient No. . | Age (yr) . | Source . | Immunocytochemistry Results . | Western Blot . | Termination of Culture . | |||
|---|---|---|---|---|---|---|---|---|
| Anticytokeratin (AE1/3+ KL-1) . | Anticytokeratin (MNF116) . | Antiepithelial (Ber-Ep4) . | Anticalretinin (08-1211) . | erbB2 Protein (21N) . | ||||
| 1 | 68 | Ascites | + | + | + | − | High | d 80 |
| 2 | 61 | Ascites | + | ND | + | ND | Int | d 90 |
| 3 | 48 | Pleura | + | + | + | − | High* | d 60 |
| 4 | 84 | Pleura | + | + | + | − | Int | d 80 |
| 5 | 49 | Ascites | + | + | + | − | High | d 80 |
| 6 | 73 | Pleura | + | + | + | − | No/low | d 130 |
Malignant effusions from breast cancer patients were cultured in α-MEM/10% FCS. The antibodies used for immunostaining and Western blot analyses are indicated in parentheses. ErbB2-protein expression was quantified by comparing the signal intensity of the erbB2-specific band (p185) with the erbB2 signals of a comparable number of either A431 or SKBR3 cells. Western blot analyses were performed on day 7 (no. 2 and 4), day 20 (no. 1 and 5), day 0/day 40 (no. 6), or day 0 (*no. 3) of culture.
Abbreviations: High, erbB2 expression comparable to SKBR3 cells; i/m, erbB2 expression higher than, or comparable to A431 cells; low, erbB2 expression lower than A431 cells; no, no detectable erbB2 expression; +/−, >95% of cells stained positive/negative; ND, not determined.