A Colony-Forming Assay for Experimental Tumours Using the Plasma Clot Diffusion Chamber
A direct tumor-colony assay in vivo was developed to measure population renewal and colonizing property as well as sensitivity of tumor stem cells to anti-cancer drugs. Ehrlich ascites tumor (EAT) cells with acquired resistance to daunomycin (DNM) have been cloned in plasma clot diffusion chambers (PCDC). Cultures were implanted into the peritoneal cavity of preirradiated mice under standard culture conditions to investigate colony growth and single cell recovery after treatment with proteolytic enzymes. The number of tumor colonies was proportional to the number of cells plated between concentrations of 0.5 x 103 to 5 x 103 cells/PCDC in resistant and non-resistant cell lines, whereas the plating efficiency appeared to be significantly lower in the resistant populations. The clones responded differently to the cytotoxic effects of DNM, with a good correlation with optimal dose schedule. This assay sufficiently warrants large scale testing of new agents to estimate the absolute number or percent of viable cancer cells after treatment.
KeywordsTumor Coloni Population Doubling Time Tumor Stem Cell Ehrlich Ascites Tumor Cell Diffusion Chamber
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