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Characterization of Cells Obtained by Mechanical and Enzymatic Means from Human Melanoma, Sarcoma, and Lung Tumors¹

Departments of Experimental Therapeutics and Grace Cancer Drug Center [H. K. S., Y. M. R., Z. P. P., W. R. G.], and Departments of Clinical Pharmacology and Therapeutics [P. J. C., Y. M. R.], Thoracic Surgery [H. T.], and Surgical Oncology [C. K.], and Computer Center [W. R. G.], Roswell Park Memorial Institute, New York State Department of Health, Buffalo, New York 14263

Characterization of cells comprising solid tumors will facilitate the rational design of cancer chemotherapy for individual patients. We have prepared cell suspensions from human melanoma, sarcoma, and lung tumors by thinly slicing the tissue with a microtome and scalpels (mechanical release), followed by treatment with a mixture of collagenase II and DNase I (enzymatic release). This method of disaggregation resulted in two cell suspensions for each tumor specimen, and we characterized these suspensions by assessing their dye exclusion capability, ribonucleoside triphosphate pools, cytological profile, and clonogenicity in soft agar. Differences between the two types of suspension and among the disease types were examined with a two-way analysis of variance. Cells released enzymatically from all disease types were characterized by a significantly higher dye exclusion capability and significantly higher pools of adenosine triphosphate and guanosine triphosphate. The two types of suspension were not significantly different in cytological profile or clonogenicity in soft agar. The enzymatic method thus yields cells in addition to those obtainable by a mild mechanical procedure, and these cells are similar in cytological profile and clonogenicity in soft agar to those released mechanically. Furthermore, the enzymatically released population is superior to that released mechanically for purposes requiring large numbers of dye-excluding cells having intact ribonucleotide pools.

¹ Supported in part by Program Grant CA-21071 and Core Grant CA-24538 from the National Cancer Institute, USPHS.

² To whom requests for reprints should be addressed, at Department of Experimental Therapeutics and Grace Cancer Drug Center, Roswell Park Memorial Institute, 666 Elm St., Buffalo, N. Y. 14263.

Received 9/ 2/80. Accepted 1/13/81.

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