Open AccessEnhanced assessment of DNA/proliferative index by depletion of tumor infiltrating leukocytes prior to monoclonal antibody gated analysis of tumor cell DNA
Author(s) -
Kenyon Norma S.,
Schmittling Robert J.,
Siiman Olavi,
Burshteyn Alexander,
Bolton Wade E.
Publication year - 1994
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990160212
Subject(s) - monoclonal antibody , flow cytometry , microbiology and biotechnology , biology , antibody , cell , cytometry , pathology , breast carcinoma , antigen , cancer research , immunology , breast cancer , cancer , medicine , genetics
Flow cytometry has become an important tool for the analysis of breast tumors, and assessment of S phase fraction and DNA ploidy are potential indicators of tumor aggression. Due to masking or dilution of infrequent tumor cell events, the presence of normal cell types, such as inflammatory cells and fibroblasts, can interfere with accurate DNA analysis of solid tumor samples. MDA‐MB‐175‐VII human breast carcinoma cells, WI‐38 human lung fibroblast cells, and peripheral blood leukocytes were mixed, in varying proportions, in order to represent human breast tumor samples. The cells were subsequently treated with CD45 conjugated magnetic microspheres to deplete tumor infiltrating leukocytes, thus enriching for tumor cells. The tumor cell mixtures were then stained with a pancytokeratin specific monoclonal antibody or with a monoclonal antibody that reacts with breast epithelial membrane antigen (EMA). When used in combination with monoclonal antibody gating, utilization of this bead‐based technology resulted in enhanced precision of DNA analysis. © 1994 Wiley‐Liss, Inc.