Detection of tumor‐specific cytotoxic drug activity IN VITRO using the fluorometric microculture cytotoxicity assay and primary cultures of tumor cells from patients

Abstract

The semi-automated fluorometric microculture cytotoxicity assay (FMCA), based on the measurement of fluorescence generated from cellular hydrolysis of fluorescein diacetate (FDA) by viable cells, was employed for cytotoxic drug sensitivity testing of tumor cells from patients with hematological or solid tumors. In total, 390 samples from 20 diagnoses were tested with up to 12 standard cytotoxic drugs. The technical success rate for different tumor types ranged from 67 to 95%. Fluorescence was linearly related to cell number but variably steep depending on tumor type. Samples from most solid tumors thus showed higher signal-to-noise ratios than hematological samples. A wide spectrum of in vitro drug activity was obtained, with acute leukemias and non-Hodgkin's lymphomas being sensitive to almost all tested drugs, whereas renal and adrenocortical carcinomas were essentially totally resistant. Between these extremes were samples of breast and ovarian carcinomas and sarcomas. When in vitro response was compared with known clinical response patterns, a good correspondence was observed. The results indicate that the FMCA is a rapid and efficient method for in vitro measurement of tumor-specific drug activity both in hematological and in solid tumors. The assay may be suitable for new drug development and direction of phase-2 trials to suitable patients.

Differential activity of cremophor EL and paclitaxel in patients' tumor cells and human carcinoma cell lines in vitro

“…Delta, i.e., the log IC50 of a cell line minus the mean log lines, U937-Vcr and 8226/Dox40, were among the cell Conversely, the melphalan-and teniposide-resis-tant 8226/LR-5 and CEM/VM-1 sublines, respectively, from patients. 7 Taxol and CEL similarity was also confirmed in the cross-resistance analysis (Table 4). Tuwere not cross-resistant to paclitaxel, indicating that increased glutathione-transferase activity and altered bulin is one potential cytoplasmic target for anticancer drugs, but the low correlation between Taxol or CEL topoisomerase II do not alter paclitaxel sensitivity.…”

“…Ongoing clinical trials using paclitaxel formulated in liposomes 30 may come to eluactivity pattern obtained in vitro is reminiscent of that in the clinic. In the authors' previous study, 7 the recidate the clinical importance of CEL. The preclinical development of Taxol relied on sponse rate to Taxol for ovarian carcinoma was considerably higher, 43%, compared with 11% in the curvarious cell lines used in vitro or in vivo as targets for the cytotoxic effect.…”

A cleavable adapter to reduce nonspecific cytotoxicity of recombinant immunotoxins

“…Total RNA was isolated using Qiagen (Hilden, Germany) midi kit according to the manufacturer's recommendations. Reverse transcription was performed using 200 U SuperScript II RT, 5 g RNA and 100 pmol oligo dT 18 . Subsequent PCR was performed using the primer set: 5Ј-GGG AAT AGT GAC TCT GAA TGT CCC CTG-3Ј and 5Ј-GAA TTC TTA GCG CAG TTC CCA CCA CTT CAG-3Ј and the product ligated into SmaI / EcoRI sites.…”

The cytotoxic activity of Taxol in primary cultures of tumour cells from patients is partly mediated by Cremophor EL

“…The present study was undertaken to investigate in vitro the activity of standard cytotoxic drugs in comparison with various formulations of paclitaxel and its solvent in tumour samples from patients and in a cell line, using the fluorometric microculture cytotoxicity assay (FMCA;Nygren et al, 1992Nygren et al, , 1994a.…”