Separation of Cells From a Fibrosarcoma in Renografin Density Gradients2

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Abstract
Cells from a murine fibrosarcoma were separated in continuous density gradients of Renografin. Four populations of viable cells appeared at densities of ≤1.064, 1.097, 1.132, and 1.170 g/cm3. The clonogenicity of these cells was determined by injection of a known number of cells intravenously into S-10-week-old mice. After 14 days, some of these cells proliferated in the lungs and appeared as macroscopic tumor nodules. The most clonogenic cells were at densities between 1.06 and 1.13. Cells banding at the heavier densities were smaller in volume and were contaminated with blood cells and/or cellular debris. The recovery of the cells was 65-85%. DNA-synthesizing cells were identified by injection of tumor-bearing mice with tritiated thymidine 1 hour before they were killed. After centrifugation, both the cell number and the acid-insoluble tritium activity were measured in each fraction of the gradient, and no significant variation in the radioactivity/cell was observed.-

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