Evaluation of a rosetting method in detection of breast cancer cells

Abstract

Epithelial membrane antigen (EMA) (Heyderman et al., 1979) is a membrane component expressed by most epithelial cells, by all breast carcinomas and their metastases but not by the vast majority of normal bone marrow cells (Sloane et al., 1980 (Goding, 1976). Briefly a 1% solution of chromium chloride (CrCl3) in 0.9% sodium chloride (NaCl) was adjusted to pH 5 by the addition of 1 N sodium hydroxide twice-weekly for 3 weeks, then stored without further adjustment in a dark container. A 0.1% solution in 0.9% NaCl was prepared freshly from this stock just prior to use. Erythrocytes from fresh heparinised blood taken from healthy volunteers were washed 4 x in 0.9% saline, and then pelleted. An aliquot of 85 p1 of the packed RBC pellet was then resuspended in a suitable volume of 0.9% saline so that when the SaM and the CrCl3 were added the resulting volume was 2ml. The antibody used was a heteroantiserum sheep-anti-mouse-immunoglobulin (SaM) dialysed extensively against 0.9% NaCl. An aliquot (0.5mg) of this antibody was added and followed immediately by 135pl of fresh 0.1% CrCl3. After 5 min at room temperature with occasional agitation, the reaction was quenched with 3-5 ml of PBS, and red cells were washed twice in PBS and resuspended in 5ml of medium. The red cells now coupled to SaM (RBC.SaM) were stored on ice overnight for use within 24 h.

11. Breast Cancer

“…With the failure of physical methods in this context, atten tion has been directed to the utility of bio chemical markers measured in the blood and/or urine [3].…”

“…These include hormones, placental proteins, enzymes and isoenzymes, antigens such as carcinoembryonic antigen (CEA), differentiated breast products such as milk proteins, breast cyst fluid and cell break down products [3]. With the monoclonal an tibody revolution, a variety of new mole cules has been detected in breast carcino mas; some of these have also been recently examined [4], While there is little doubt that biochemi cal indices may be elevated in patients with end-stage metastatic disease, their real utility would be in forecasting the evolution of mé tastasés undetected by other means.…”