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Large-Capacity Separation of Malignant Cells and Lymphocytes from the Furth Mast Cell Tumor in a Reorienting Zonal Rotor¹

Departments of Pathology [C. L. G., T. P. P., K. A. T., W. J. C., A. M. P., T. G. P.], Biochemistry [T. G. P.], and Biostatistics [E. L. B.], University of Alabama Medical Center, Birmingham, Alabama 35294

The Furth murine mastocytoma was adapted to the ascitic form and separated into fractions enriched with respect to lymphocytes and malignant cells by velocity sedimentation in the SZ-14 reorienting zonal rotor or in the isokinetic gradient. Lymphocytes were more highly purified (p < 0.01) in the isokinetic gradient than in the zonal rotor, i.e., lymphocytes comprised 99.1% of the nucleated cells in the purest fraction from the isokinetic gradient and 80.1% of the nucleated cells in the purest fraction from the zonal rotor. Neoplastic mast cells were similarly purified by the two methods; they comprised 67.7 and 78.5% of the nucleated cells in the purest fractions from the isokinetic gradient and zonal rotor, respectively. Up to 160 million tumor cells can be purified in a single step with the reorienting zonal rotor, whereas 30 to 40 million cells per gradient approach the limit of the isokinetic gradient. After centrifugation in the zonal rotor, recovery was 85.6 ± 12% (S.D.) of the cells layered over the gradient; and the separated tumor cells retained their ability to form tumors when transplanted into mice. The separation of large numbers of lymphocytes and malignant cells from the same tumor in the SZ-14 rotor should aid in the biochemical and immunological characterization of cancer.

¹ This research was supported by USPHS Grants CA-13148 and CA-23922 from the National Cancer Institute and by American Cancer Society Grant PDT-126.

² To whom requests for reprints should be addressed, at Departments of Pathology and Biochemistry, University of Alabama Medical Center, University Station, Birmingham, Alabama 35294.

Received 9/ 4/79. Accepted 2/27/80.