Production of Viable Single Ceil Suspensions From Solid Tumors

A method is described for utilizing trypsin and DNAase to obtain suspensions of large numbers of single viable cells from a variety of solid tumor sources. Representative yields are reported. The viability index, which was obtained by vital staining, approached 100 percent. Metabolic studies disclosed little or no alteration in respiration and aerobic and anaerobic glycolysis of enzyme-processed cells, whether they were derived from solid tumors or from ascites cells and compared to unprocessed ascites cells. Processed cells derived from solid tumors showed consistently higher metabolic values than tissue slices of the tumor. Testosterone inhibited, and 2,4-dinitrophenol and endotoxin stimulated, certain metabolic parameters in tumor cells where such responses could be expected. The cells from one of two solid tumors tested compared favorably with equal numbers of the same strain ascites cells in ability to form tumors when injected back into mice of the host strain. Few, if any, morphologic differences were seen microscopically between stained cells derived from solid tumors and those from the ascites counterpart. Cells processed from a solid tumor were maintained in continuous tissue culture for 3 months. Time-lapse photography disclosed amoeboid motion. After 6 days in culture, biological virulence comparable to that in freshly derived cells was demonstrated by injection back into mice of the host strain.