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A Nonclonogenic Cytotoxicity Assay Using Primary Cultures of Patient Tumor Cells for Anticancer Drug Screening

Division of Clinical Pharmacology, University Hospital, Uppsala University, Uppsala, Sweden

Joachim Gullbo

Division of Clinical Pharmacology, University Hospital, Uppsala University, Uppsala, Sweden

Kenneth Nilsson

Department of Pathology, University Hospital, Uppsala University, Uppsala, Sweden

Peter Nygren

Department of Oncology, University Hospital, Uppsala University, Uppsala, Sweden

Rolf Larsson

Division of Clinical Pharmacology, University Hospital, Uppsala University, Uppsala, Sweden

Primary cultures of cells from patients with chronic lymphocytic leukemia (CLL) and ovarian carcinoma (Ovca) were compared with the renal carcinoma ACHN and the lymphocytic CCRF-CEM human tumor cell lines in response to 63 toxic or nontoxic compounds. The experiments were conducted at 1, 10, and 100 µ/ml in 96-well microtiter plates for an assay time of 72 h. The plates were analyzed by the fully automated Dynatech Immuno Assay System (DIAS) using Alamar blue as a fluorometric/colorimetric indicator of metabolic activity. Drug response data were reported as the area under the tumor cell survival-concentration curve (AUC). Noncytotoxic compounds were classified as inactive by all cell systems. According to the AUC, CCRF-CEM and CLL cells were the most sensitive, followed by ACHN, and then Ovca. Many of the clinically active drugs were detected by all cell systems. However, the sensitivity pattern differed considerably between the cell types as judged from correlation analysis, and a higher proportion of clinically inactive drugs were scored as active by the cell lines compared with the primary cultures. Ovca showed the highest ratio of clinically solid tumor active/clinically inactive agents followed by CLL. The results indicate that primary cultures of human tumor cells may be a useful and valuable model for anticancer drug screening.

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