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Separation of Cells from Mouse Solid Tumors by Centrifugal Elutriation¹

Section of Experimental Radiotherapy [M. L. M., D. J. G.], Department of Physics [R. E. M.], and Department of Developmental Therapeutics [B. B.], The University of Texas System Cancer Center, M. D. Anderson Hospital and Tumor Institute, Houston, Texas 77030

Centrifugal elutriation was used to separate cells dissociated from two hypotetraploid mouse solid tumors, a fibrosarcoma and a sarcoma derived from L-P59 cells, based on their sedimentation rates. The separation was rapid, requiring less than 1 hr; yielded about 80% cell recovery; and resulted in little loss of cell viability. Analysis of DNA content by flow cytometry demonstrated the synchrony obtained with these tumor cells. The fractions with the lowest sedimentation rates contained predominantly, normal cells, those with intermediate sedimentation rates contained predominantly tumor cells in the G₁ phase of the cell cycle, and those with the highest sedimentation rates contained mostly tumor cells in S or G₂. The clonogenicity of L-P59 cells, assayed in culture, markedly increased with increasing sedimentation rates. In contrast, the clonogenicity of fibrosarcoma cells, assayed in vivo by a lung colony assay, was lower for the smaller cells, but was essentially constant among the larger cells. Autoradiography of cells labeled in vivo with tritiated thymidine demonstrated no differences in the proportions of cycling cells in various fractions. These results demonstrate that subpopulations differing in cell type, phase of the cell cycle, and clonogenicity can be rapidly separated from solid tumors by centrifugal elutriation.

¹ This study was supported in part by the Department of Health, Education and Welfare, NIH, National Cancer Institute Grants CA-06294, CA-17364, CA-04484, CA-18628, CA-11430, and CA-14528.

Received 6/29/77. Accepted 9/ 6/77.

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