Transformation of BALB/c-3T3 Cells: I. Investigation of Experimental Parameters That Influence Detection of Spontaneous Transformation

Matthews, Edwin J.

doi:10.2307/3431401

Cited by 6 publications

(17 citation statements)

References 16 publications

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“…The investigations in this report used the 1-13 clone of A31 BALB/c-3T3 cells (15,16). The materials and methods used to culture the cells have been previously reported in detail (2) and are summarized in part I of these investigations (17).…”

Section: Cell Culturementioning

confidence: 99%

Transformation of BALB/c-3T3 cells: III. Development of a co-culture clonal survival assay for quantification of chemical cytotoxicity in high-density cell cultures.

Matthews

1993

Environ Health Perspect

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A co-culture clonal survival assay was developed to measure the cytotoxicity of test chemical treatments to BALB/c-3T3 cells because the standard clonal survival assay using 200 wild type (WT) cells frequently overestimates chemical cytotoxicity when compared with identical treatment doses in high-density cultures. The assay used co-cultures of 3.2 x 10(4) WT cells, the same seeding density used in the transformation assay, and 200 ouabain resistant (OUAr) cells. After a 48-hr test chemical treatment, co-cultured cells were fed with culture medium containing 4 mM ouabain. The test chemical was cytotoxic to an equal percentage of WT and OUAr cells. Ouabain treatments killed the remaining WT cells. Thus, OUAr cells surviving the test chemical treatment measured the relative cloning efficiency (RCE) of all treated cells in the high-density cell co-culture. The co-culture assay succeeded because metabolic cooperation at the OUAr locus was not detected in BALB/c-3T3 cells. Five chemicals induced comparable cytotoxic responses in both assays, including actinomycin D, 5-bromo-2'-deoxyuridine, N'-methyl-N-nitro-N'-nitrosoguanidine, dimethyl sulfoxide and sodium chloride. In contrast, chemical cytotoxic responses detected in the standard and co-culture assays differed by > or = 10-fold for 11-aminoundecanoic acid, benzo[a]pyrene, cytosine arabinoside, and 3-methyl-cholanthrene and differed by > 2-fold for 2-acetylaminofluorene and dimethylnitrosamine. Detection of 11-aminoundecanoic acid-induced transformation was shown to be dependent on selecting treatment doses from the co-culture assay data. Thus, this method permits more accurate assessment of both chemical-induced cytotoxicity and transformation.

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Section: Cell Culturementioning

confidence: 99%

Transformation of BALB/c-3T3 cells: III. Development of a co-culture clonal survival assay for quantification of chemical cytotoxicity in high-density cell cultures.

Matthews

1993

Environ Health Perspect

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“…A BALB/c 3T3 cell transformation assay was performed according to a modified protocol as previously described [22]–[26]. Cells were seeded at a density of 10 4 cells/60 mm dish and then incubated for 24 hours.…”

Section: Methodsmentioning

confidence: 99%

Detection of Cell Carcinogenic Transformation by a Quadruplex DNA Binding Fluorescent Probe

et al. 2014

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Cancer can be easily treated when found early. A probe capable of detecting cell transformation may increase the success rate of early diagnosis of cancer. In this report we have tested the ability of a fluorescent, quadruplex DNA binding probe, 3,6-bis(1-methyl-4- vinylpyridinium) carbazole diiodide (BMVC), to detect cell transformation in vitro. BMVC was applied to living cells in several different models of cell transformation, and the fluorescence signals of BMVC were measured. The degrees of cell transformation in these models were characterized by alterations in cellular morphological phenotype and subcellular organization. When BMVC probes were applied, the number of BMVC-positive cells increased in accordance with the degree of transformation. BMVC was capable of significantly detecting formation of foci, increased cellular motility, cell proliferation, cell apoptosis, anchorage-independent growth, and increased invasiveness of transformed cells. These results demonstrate the ability of BMVC probes to detect cell transformation and indicate that BMVC is of promise for use as a probe in early cancer detection.

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“…In contrast, the cell transformation assay was positive in one study and negative in another. The significance of the positive finding is equivocal, as cell transformation has a poor record of reproducibility among laboratories, and is very sensitive to such variables as the source of foetal bovine serum in the growth medium, the passage number and the source of cryopreserved cells (36). Although inorganic Mn has been shown to reduce the fidelity of DNA replication in pure in vitro cell-free systems, postulated to be due to interaction with the active sites of DNA polymerase (10,15), the evidence for metal mutagenicity in more conventional genotoxicity assays is inconsistent (49) and difficult to assess in relation to human risk.…”

Section: Genetic Toxicologymentioning

confidence: 99%

General toxicology of MnDPDP

Larsen¹,

Grant²

1997

Acta Radiol

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Purpose:To investigate the general toxicology of mangafodipir trisodium (Mn-DPDP, Teslascan).Material and Methods: Studies were performed in accordance with standard methods and in compliance with regulations current at the time of conduct.Results: Single-dose studies in rodents and dogs showed that MnDPDP was tolerated at doses of approximately 2000 ymolkg, approximately 400 times a single imaging dose of 5 pmoVkg. The single dose tolerance of MnDPDP was approximately 10 times greater than MnC1,. A good safety profile of MnDPDP was also shown in repeat-dose studies (3 weeks), in which the no-observed-adverse-effect level for the rat, monkey and dog was 116,29 and 10 ymoVkg, respectively. The local tolerance studies indicated that no adverse local tissue reactions are likely to occur after i.v. injection. Other studies indicate that accidental spillage of MnDPDP onto the skin is not expected to lead to significant systemic exposure, or to local irritation or hypersensitivity. MnDPDP was not genotoxic in a battery of several different tests.Conclusion: MnDPDP was shown to have a good safety profile suitable as an hepatobiliary MR contrast agent for i.v. administration. ommended daily allowance for adults of 2-5 mg (46). The purpose of this paper is to summarize several of the general toxicology studies on MnDPDP, including single and repeat-dose studies, genetic toxicology, local tolerance, antigenicity and skin permeability which, together with other pre-clinical and clinical data (17,28,31,33,45,62,64,67), aims to provide a sufficiently detailed toxicological profile of MnDPDP for an objective assessment of safety.

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Transformation of BALB/c-3T3 Cells: I. Investigation of Experimental Parameters That Influence Detection of Spontaneous Transformation

Cited by 6 publications

References 16 publications

Transformation of BALB/c-3T3 cells: III. Development of a co-culture clonal survival assay for quantification of chemical cytotoxicity in high-density cell cultures.

Transformation of BALB/c-3T3 cells: III. Development of a co-culture clonal survival assay for quantification of chemical cytotoxicity in high-density cell cultures.

Detection of Cell Carcinogenic Transformation by a Quadruplex DNA Binding Fluorescent Probe

General toxicology of MnDPDP

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