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Simple Method for Quantifying Pulmonary Metastases in a Murine Fibrosarcoma

Newport Pharmaceuticals International, Inc., Newport Beach, California 92660-0147 [C. E. J., C. T. L.]; and The Department of Immunopathology, The Cleveland Clinic Foundation, Cleveland, Ohio 44106 [S. D. D.]

A simple method is described for quantitating pulmonary metastases in C57BL/6J mice implanted with the Lewis T241 fibrosarcoma. Lungs were removed from tumor-bearing mice, homogenized, and diluted to 14 ml with supplemented Roswell Park Memorial Institute Tissue Culture Medium 1640 (+5% fetal calf serum). Twenty-five µl of lung suspension from individual mice were added to a flat-bottomed microtiter plate containing 175 µl of supplemented media, pulsed with [³H]thymidine, and incubated for 20 hr at 37°. The cells were processed with a multiple automated sample harvester and counted in a liquid scintillator. Total lung burden was extrapolated from measured radioactivity (as cpm) through the use of a standard curve. This method is reasonably accurate and precise and provides more quantitative information on the pulmonary metastatic process than most conventional assays.

¹ To whom requests for reprints should be addressed, at Newport Pharmaceuticals International, Inc., 897 West 16th Street, Newport Beach, Calif. 92660-0147.

Received 3/28/83. Accepted 8/19/83.