A NOVEL DYE EXCLUSION METHOD FOR TESTING INVITRO CHEMOSENSITIVITY OF HUMAN-TUMORS

  • 1 January 1983
    • journal article
    • research article
    • Vol. 43  (2) , 749-757
Abstract
Dissociated cancer cells are exposed to antineoplastic drugs (5 .times. 104 viable cells/drug for 1 h or continuously) and cultured for 4-6 days in liquid medium. Cells are then stained with Fast green dye, sedimented onto slides with a Cytospin centrifuge, and counterstained with a modified hematoxylin and eosin technique. Dead cells stain with Fast green, and living cells stain with hematoxylin and eosin. Cell kill is calculated as percentage of control based on the relative numbers of living tumor cells, living non-tumor cells and dead cells. Drug sensitivity could be assayed in 125 of 162 specimens of human neoplasms obtained from malignant effusions (16 of 18), excisional biopsies (31 of 44), needle biopsies (34 of 47), endoscopic biopsies (18 of 23), peripheral blood samples (19 of 20) and bone marrow aspirates (5 of 7). The assays were successful (median of 10 drugs [Doxorubicin, nitrogen mustard, melphalan, carmustine, cis-platin, mitomycin C, vinblastine, 5-fluorouracil, UP-16 (etoposide) and dexamethasone] tested) in: 46 of 64 andenocarcinomas; 4 of 11 squamous cell carcinomas; 5 of 7 lymphomas; 6 of 7 melanomas; 2 of 4 sarcomas; 18 of 20 transitional-cell carcinomas; 18 of 20 transitional-cell carcinomas; 14 of 15 small-cell carcinomas; 7 of 8 myelomas; 12 of 12 chronic lymphocytic leukemias; 7 of 9 acute leukemias and 4 of 5 undifferentiated carcinomas. The assay results demonstrated a strong correlation between the in vitro chemosensitivity of different types of tumors and the known clinical response patterns of these tumors. This assay can be used to determine which specific cells are killed in a heterogeneous cell population. Further work is needed to determine if this assay may be useful in blind screening trials for antineoplastic agents or if it may be of clinical use in predicting response to agents which are not cycle specific.