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A rapid technique for isolation of viable tumor cells from solid tumors: Use of the tumor cells for induction and measurement of cell-mediated cytotoxic responses

1982; Pergamon Press; Volume: 18; Issue: 10 Linguagem: Inglês

10.1016/0277-5379(82)90248-6

1878-5980

Eli Kedar, Barbara Ikejiri, Guy D. Bonnard, Ronald B. Herberman,

Immunotherapy and Immune Responses

A rapid and simple technique for the isolation of viable tumor cells from human and mouse solid neoplasms is described. It consists of a 5 to 10-min treatment with trypsin-collagenase-DNase mixture, followed by mechanical disaggregation of the tumor tissue and subsequently by a brief centrifugation on a discontinuous Percoll gradient. With the tumors employed, this procedure usually requires less than 1 hr and results in preparations comprising greater than 80% tumor cells with viability of 80–90%. Cell-mediated cytotoxic response was measured with: (a) unsensitized lymphocytes freshly obtained from tumor-bearing hosts; (b) lymphocytes propagated in culture with T cell growth factor; and (c) lymphocytes stimulated in cocultures with autologous or syngeneic tumor cells. The cytotoxic activity was assessed in a modified [51Cr]-release assay adapted for solid tumor cells, allowing a long incubation period (24 hr) and the use of a low number (200–1000) of highly labeled target cells (2–10 counts/min/cell).