In Vitro Transformation Assays Using Mouse Embryo Cell Lines

Rundell, John O.

doi:10.1007/978-1-4612-5164-4_20

Cited by 10 publications

(17 citation statements)

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“…Appendix A. 59 (15) 110 (15) 91 (15) 32 (15) 40 (30) 152 (20) 115 (20 127 (20) 75 (20) 6 (12,20) 4 (9,20) 7 (20) 24 (20) 24 (20) 150 (20) 126 (20) 6. 14** (40) 128 (20) 71 (20) .…”

Section: Discussionunclassified

“…59 (15) 110 (15) 91 (15) 32 (15) 40 (30) 152 (20) 115 (20 127 (20) 75 (20) 6 (12,20) 4 (9,20) 7 (20) 24 (20) 24 (20) 150 (20) 126 (20) 6. 14** (40) 128 (20) 71 (20) . (20) 15 (20) 31 (20) 22 (40) 128 (20) 71 (20) 21 ( 33 (20) 36 (20) 18 (20) 17 (20) (8,18) 72 (18) 62 (18) 39 (18) 150 (71) .189 2.97* 3.13*** 1.90 1.52 6 (20) 5 (20) 3 (19) 4 (20) 31 (75) 59 (20) 40 (19) 0 (4,20) 0 (20) 10 (20) 9 (20) 58 (77) .…”

Section: Discussionunclassified

“…14** (40) 128 (20) 71 (20) . (20) 15 (20) 31 (20) 22 (40) 128 (20) 71 (20) 21 ( 33 (20) 36 (20) 18 (20) 17 (20) (8,18) 72 (18) 62 (18) 39 (18) 150 (71) .189 2.97* 3.13*** 1.90 1.52 6 (20) 5 (20) 3 (19) 4 (20) 31 (75) 59 (20) 40 (19) 0 (4,20) 0 (20) 10 (20) 9 (20) 58 (77) . .000…”

Section: Discussionunclassified

“…16 (20) 30 (20) 43 (19) 35 (20) 89 (78) 60 (20) 20 (20) (20) 34 (19) 29 (20) 29 (40) 90 (20) 24 (20) 21 (20) 4 (20) 2 (18) …”

Section: Discussionunclassified

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Transformation of BALB/c-3T3 cells: IV. Rank-ordered potency of 24 chemical responses detected in a sensitive new assay procedure.

Matthews

Spalding

Tennant

1993

Environ Health Perspect

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This report introduces an improved method of detecting chemical-induced morphological transformation of A-31-1-13 BALB/c-3T3 cells. The new procedure uses an increased target cell population to assess chemicalinduced damage by increasing the initial seeding density and by delaying the initiation time of chemical treatment. Furthermore, a newly developed co-culture clonal survival assay was used to select chemical doses for the transformation assay. This assay measured the relative cloning efficiency (RCE) of chemical treatments in high-density cell cultures. In addition, transformation assay sensitivity was enhanced through the use of improved methods to solubilize many chemicals. From a group of 24 chemicals tested in at least two trials, clear evidence of chemical-induced transformation was detected for 12 chemicals (aphidicolin, barium chloride-2H20, 5-bromo-2'-deoxyuridine, C.I. direct blue 15, trans-cinnamaldehyde, cytosine arabinoside, diphenylnitrosamine, manganese sulfate-H20, 2-mercaptobenzimidazole, mezerein, riddelliine, and 2,6-xylidine); 2 chemicals had equivocal activity [C.I. direct blue 218 and mono(2-ethylhexyl)phthalate], 9 chemicals were inactive [carisoprodol, chloramphenicol sodium succinate, 4-chloro-2-nitroaniline, C.I. acid red 114, isobutyraldehyde, mono(2-ethylhexyl)adipate, sodium fluoride, and 12-O-tetradecanoylphorbol-13-acetate), and 1 chemical had an indeterminate response (2,6-dinitrotoluene). All positive responses were detected in the absence of an exogenous activation system and exhibited significant activity at two or more consecutive doses. This report also presents a mathematical method that uses t-statistics for rank-ordering the potency of chemical-induced transformation responses. This model detects sensitivity differences in experiments used to evaluate chemical-induced transformation. Furthermore, it provides a method to estimate a chemical's transformation response in terms of the historical behavior of the assay, as well as its future activity. The most active of the 24 chemicals was mezerein, and the least active chemical was diphenylnitrosamine.

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“…Appendix A. 59 (15) 110 (15) 91 (15) 32 (15) 40 (30) 152 (20) 115 (20 127 (20) 75 (20) 6 (12,20) 4 (9,20) 7 (20) 24 (20) 24 (20) 150 (20) 126 (20) 6. 14** (40) 128 (20) 71 (20) .…”

Section: Discussionunclassified

“…16 (20) 30 (20) 43 (19) 35 (20) 89 (78) 60 (20) 20 (20) (20) 34 (19) 29 (20) 29 (40) 90 (20) 24 (20) 21 (20) 4 (20) 2 (18) …”

Section: Discussionunclassified

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Transformation of BALB/c-3T3 cells: IV. Rank-ordered potency of 24 chemical responses detected in a sensitive new assay procedure.

Matthews

Spalding

Tennant

1993

Environ Health Perspect

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“…Briefly, the number of type I-III transformed foci of BALB/c-3T3 cells were identified microscopically using published criteria (5)(6)14,(18)(19), and type III foci had three phenotypic properties: piling and overlapping cells, disorientation of cells at the periphery of the focus, and invasion of transformed cells into a contact-inhibited monolayer of WT cells. Type I and II foci also appeared in many different sizes, but they lacked one or more of the three phenotypic properties of the type III transformed focus.…”

Section: Evaluation Of Transformed Focimentioning

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Transformation of BALB/c-3T3 Cells: II. Investigation of Experimental Parameters That Influence Detection of Benzo[a]pyrene-Induced Transformation

Matthews¹

1993

Environmental Health Perspectives

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Benzo[alpyrene (BaP) induced significant morphological transformation of clone A31-1-13 BALB/c-3T3 cells without exogenous activation. Therefore, BaP was selected as a model to determine the internal consistency of detection of chemical-induced transformation. BaP induced a continuum of type I-III foci of different sizes, and the ratio of type I-III to type III foci/vessel was usually about 2-fold. The major finding was that BaP induced highly significant transformation responses, and the magnitude of these responses were inversely correlated with the cytotoxicity of the treatment doses. Thus, the induction of BaP-induced transformation behaved as though it was caused by a mutational event. Variability among responses were shown to depend on the serum lot and the cryopreserved ampule of cells. In addition, experiments with low spontaneous transformation responses had an impaired ability to detect BaP; however, experiments with high or normal spontaneous responses had a normal ability to detect BaP. Because the expression of BaP-induced transformation depended on both the cytotoxicity of the treatment and the cumulative number of mitoses, the frequency of BaP-induced transformation should be reported as the number of foci/vessel, but not expressed as the number of foci/viable cell surviving the chemical treatment. These conclusions are important because the same 110 experiments described in this report were also used to evaluate the transformation responses of many different carcinogenic and noncarcinogenic chemicals. These data are being reported separately.

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Assessment of chemical carcinogen-induced transforming activity using BALB/c-3T3 cells

Matthews¹

1986

Journal of Tissue Culture Methods

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SUMMARY:The BALB/c-3T3 cell transformation assay evaluates the morphologic transforming potential of test chemicals using cultured mammalian cells as targets. The assay is semiquantitative and employs a contactinhibited clone of cells derived from the original BALB/c-3T3 mouse cell line. Transforming activity, or a focus, is recognized as a dense layer of morphologically altered cells superimposed on a monolayer of the normal cell phenotype. The induction of transforming activity in a standard assay has been reported to correlate well with the carcinogenicity of many test chemicals in rodent bioassay; however, the standard assay does not detect carcinogens of all chemical classes. This report describes an operational protocol for the standard BALB/c-3T3 cell transformation assay. In addition, it provides assay acceptance criteria and a discussion of a method to evaluate transforming activity data.

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In Vitro Transformation Assays Using Mouse Embryo Cell Lines

Cited by 10 publications

References 4 publications

Transformation of BALB/c-3T3 cells: IV. Rank-ordered potency of 24 chemical responses detected in a sensitive new assay procedure.

Transformation of BALB/c-3T3 cells: IV. Rank-ordered potency of 24 chemical responses detected in a sensitive new assay procedure.

Transformation of BALB/c-3T3 Cells: II. Investigation of Experimental Parameters That Influence Detection of Benzo[a]pyrene-Induced Transformation

Assessment of chemical carcinogen-induced transforming activity using BALB/c-3T3 cells

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