Development of a 9L Rat Brain Tumor Cell Multicellular Spheroid System and Its Response to 1,3-Bis(2-chloroethyl)-1-nitrosourea and Radiation2

Deen, Dennis F.; Hoshino, Takao; Williams, Mary Elizabeth; Muraoka, Isao; Knebel, Kathy D.; Barker, Marvin

doi:10.1093/jnci/64.6.1373

Cited by 48 publications

(20 citation statements)

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“…Both panels of Figure 1 show cellular radiosensitivity as a function of Hoechst staining intensity; in spheroids, the symmetry of the model allows an easy calculation of the depth from which the cells were recovered (Durand, 1983), while in the tumour system, the data are simply presented as fraction number, where fraction I represented the 10% of the cells which were most intensely stained (thus nearest the functional blood supply, e.g. Chaplin et al, 1985Chaplin et al, ,1986a (Figure 2) indicated that the less well oxygenated cells were more sensitive to this nitrosourea, in agreement with previous results obtained with nitrosoureas in other spheroid systems (Deen et al, 1980;Kwok & Twentyman, 1985). Quite reproducible responses were found in the multiple experiments shown (note that in the case of spheroids, different starting sizes led to slightly different estimates of cell depth among experiments).…”

Section: Resultssupporting

confidence: 85%

Chemosensitization by misonidazole in CCNU-treated spheroids and tumours

Durand

Dj²

1987

Br J Cancer

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Summary Misonidazole has been demonstrated to enhance the cytotoxicity of several common antineoplastic drugs in vitro and in vivo, and its mechanism of action as a chemosensitizer, though still unknown, is thought to be dependent upon hypoxia. We have used fluorescence-activated cell sorting to evaluate chemopotentiation by misonidazole as a function of cell position in V79 spheroids and KHT tumours. CCNU toxicity was enhanced in all cell subpopulations of both tumours and spheroids, with greater consistency than might be predicted on the basis of the known variations in oxygen tension. Further, both misonidazole and CCNU as single agents were preferentially toxic in the less well oxygenated regions of each system, arguing that differential toxicity canno t be implicated in the chemopotentiation observed. In fact, increased treatment toxicity did not necessarily lead to increased chemopotentiation, nor was potentiation directly related to the metabolism/binding of the misonidazole. Chemopotentiation in multicell systems thus appears to be a complex, multi-factorial process.Hypoxic cells develop spontaneously during the growth of many solid tumours, and are known to be resistant to ionizing radiation and implicated in resistance to several antineoplastic drugs. Clearly, their presence in human tumours may limit the effectiveness of cancer therapy. Development of hypoxic cell radiosensitizers thus has a clear rationale; the observations that many of these sensitizers were preferentially toxic to hypoxic cells (Olive & McCalla 1975; Hall & Roizin-Towle 1975;Moore et al., 1976) suggested that they may be complementary to many conventional cancer chemotherapeutic agents, and could thus be used effectively in combination treatments. Indeed, in a number of instances an interaction between the sensitizer and the chemotherapeutic agent was observed (Kelly et al., 1979;Clement et al., 1980;Rose et al., 1980, and reviewed by Siemann, 1984).Many factors have been implicated in the phenomenon of chemosensitization (see Brown 1982, and Siemann 1984 for reviews), including differential toxicity (Kelly et al., 1979), direct interactions between the sensitizer (or its toxic metabolites, which are generally produced under hypoxic conditions) and the chemotherapeutic agent itself (Taylor et al., 1982), alterations of drug pharmacology or delivery (Urtasun et al., 1982;Workman et al., 1983;Lee & Workman 1986), inhibition of repair processes by chemosensitizer treatment (Taylor et al., 1982;Mulcahy 1986), and alterations of cellular drug sensitivity, by, for example, the sensitizer selecting for surviving cells more vulnerable to the chemotherapeutic agent due to cell cycle status or other factors . Perhaps the most consistent observation, however, has been the apparent requirement of hypoxia both in vitro (Brown, 1982;Mulcahy, 1984), and in vivo Wheeler et al., 1984).In this report, we present data evaluating the cytotoxic effects of misonidazole, MISO, and N-(2-chloro-ethyl)-N'-cyclohexyl-N-nitrosourea, CCNU, singly and in combi...

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Section: Resultssupporting

confidence: 85%

Chemosensitization by misonidazole in CCNU-treated spheroids and tumours

Durand

Dj²

1987

Br J Cancer

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“…It has been shown that the resistance of cells in spheroids to VCR may be partly related to the limited penetration of this drug in the spheroid and partly to the "out of cycle" state of many cells in spheroids (Wibe & Oftebro, 1981). Limited penetration into spheroids has also been shown for a closely-related drug, vinblastine (Nederman, 1984 (Deen et al, 1980) and V79 cells (Hetzel & Kaufman, 1983) which showed single cells to be more resistant than cells in spheroids.…”

Section: Resultsmentioning

confidence: 86%

“…Factors related to this structure, such as intercellular contact, may be involved in determining the response of cells within the spheroid to radiation and cytotoxic drugs. (Durand & Sutherland, 1972;Sutherland et al, 1979;Deen et al, 1980;Dertinger & Hiilser, 1981;Wibe & Oftebro, 1981). The initial study by Durand & Sutherland (1972) showed that Chinese hamster V79 cells were more resistant to radiation when irradiated as aggregates than as single cells and these authors suggested that intercellular contact may be responsible for the observation.…”

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confidence: 99%

“…Received 14 September 1984;and in revised form, 24 October 1984. the extent of intercellular ionic coupling. The cells within spheroids from other cell lines have also been reported to be more resistant than single cells to treatment with hyperthermia (Durand, 1978) and cytotoxic drugs, such as adriamycin (Sutherland et al, 1979), vincristine (Wibe & Oftebro, 1981) and vinblastine (Nederman, 1984), In contrast, however, single cells were found to be more sensitive than cells within intact spheroids to BCNU (Deen et al, 1980) or nitrogen mustard (Hetzel & Kaufman, 1983).…”

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confidence: 99%

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The response to cytotoxic drugs of EMT6 cells treated either as intact or disaggregated spheroids

Kwok¹,

Twentyman²

1985

Br J Cancer

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Summary We have compared the response to a number of cytotoxic drugs of cells treated either within intact multicellular spheroids or as isolated cells following spheroid disaggregation. The cells used were of the EMT6/Ca/VJAC mouse tumour line and spheroids were treated or disaggregated at a mean diameter of 250,um. The response of cell to nitrogen mustard (HN2) or CCNU was similar under the two exposure conditions and we conclude that factors related to spheroid structure (i.e. drug penetrability, intercellular contact effect and microenvironment within the spheroid) do not influence the initial response to these agents. Recovery of potentially lethal damage occuring over 24h, however, greatly modifies the level of cell killing in intact spheroids. EMT6 cells were found to be extremely resistant to vincristine under all exposure conditions. For adriamycin (ADM), cells were always initially more sensitive when exposed to the drug in suspension rather than in intact spheroids. When ADM exposure was prolonged beyond 1 h, however, delaying spheroid disaggregation for 24h led to increased cell kill and reduced differential between the two conditions of exposure. The data suggest that both drug penetration problems and other factors related to spheroid structure are involved in determining the response of cells in small spheroids to ADM.

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“…Oxygen does appear to be available to all the cells in 400 pm diameter 9L spheroids, at least to the level required to maintain an oxic radiosensitivity, since the radiation dose response of this size spheroid has no radioresistant hypoxic fraction. 29 The finding that a substantial fraction of the cells in 400 pm diameter spheroids of the V-79 line is radioresi~tant~' emphasizes that conclusions regarding nutrient distribution in tumor system for one cell line should not be extrapolated to other cell lines.…”

Section: 8mentioning

confidence: 99%

The glucose distribution in 9l rat brain multicell tumor spheroids and its effect on cell necrosis

1982

Cancer

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The glucose consumption rate of 9L rat brain tumor cells was determined as a function of concentration. Glucose uptake followed Michaelis‐Menten kinetics with a Km of 0.58 mM and a vmax of 1.6 pg/cellmin. The glucose diffusion coefficient in spheroids of 9L tumor cells was determined to be 1.5 × 10−6 cm2/s at 37°C. Using these parametric values, the glucose distribution in 9L multicell spheroids was calculated and related to the viable and necrotic zones.

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Development of a 9L Rat Brain Tumor Cell Multicellular Spheroid System and Its Response to 1,3-Bis(2-chloroethyl)-1-nitrosourea and Radiation2

Cited by 48 publications

References 0 publications

Chemosensitization by misonidazole in CCNU-treated spheroids and tumours

Chemosensitization by misonidazole in CCNU-treated spheroids and tumours

The response to cytotoxic drugs of EMT6 cells treated either as intact or disaggregated spheroids

The glucose distribution in 9l rat brain multicell tumor spheroids and its effect on cell necrosis

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