Sarcoma-Producing Cell Lines Derived from Clones Transformed in vitro by Benzo[
            <i>a</i>
            ]pyrene

DiPaolo, Joseph A.; Nelson, Richard L.; Donovan, Paul

doi:10.1126/science.165.3896.917

Science

1969

DOI: 10.1126/science.165.3896.917

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Sarcoma-Producing Cell Lines Derived from Clones Transformed in vitro by Benzo[ a ]pyrene

Joseph A. DiPaolo

Richard L. Nelson

Paul Donovan

Abstract: Cells derived from mixed Syrian hamster embryo cultures were treated with pyrene (control) or with benzo[a]pyrene. Transformed clones were obtained only with the carcinogen. Some of the transformed clones were responsible for cell lines that produced tumors when injected into hamsters. These observations provide evidence that chemical-induced oncogenesis can be studied by an in vitro model.

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Cited by 64 publications

(33 citation statements)

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“…However, slight toxicity was observed at 5.0 and 10.0 ,ug/ml for six days and at 10.ug/ml for one day. t These cultures were observed for 120 days and then discarded.…”

mentioning

confidence: 99%

“…the toxicity the compounds exert.7 Other investigators working with cells from sources different form ours have also successfully achieved systems for obtaining chemical carcinogenesis in vitro.8- 10 We shall herewith define the word "transformation" to denote the phenotypic state in which cells form piled-up colonies that give rise to fibrosarcomas on inoculation into isologous mice.…”

mentioning

confidence: 99%

See 1 more Smart Citation

In vitro Malignant Transformation by Methylcholanthrene of the Progeny of Single Cells Derived from C3H Mouse Prostate

Mondal

Heidelberger

1970

Proc. Natl. Acad. Sci. U.S.A.

103

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Abstract. Cell lines derived from C3H mouse ventral prostate grow to a monolayer, do not pile up, and do not give tumors on inoculation into isologous mice. When treated with carcinogenic hydrocarbons, these cells produce piledup colonies that give rise to fibrosarcomas in mice; these colonies are termed transformed. Individual single cells were treated with 0.5 per cent dimethyl sulfoxide (DMSO), and 2 per cent of them gave rise to clones containing transformed colonies. Under optimal conditions, treatment of individual single cells with 3-methylcholanthrene (MCA) led to the formation of transformed colonies in 100 per cent of the clones. From recloning experiments, it appears probable that all the progeny of the MCA-treated cells were potentially transformed, although the time required for the transformation to be scored was variable. It is concluded that in this system the carcinogen does not select for pre-existing malignant cells.

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“…However, slight toxicity was observed at 5.0 and 10.0 ,ug/ml for six days and at 10.ug/ml for one day. t These cultures were observed for 120 days and then discarded.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

In vitro Malignant Transformation by Methylcholanthrene of the Progeny of Single Cells Derived from C3H Mouse Prostate

Mondal

Heidelberger

1970

Proc. Natl. Acad. Sci. U.S.A.

103

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“…The ability of chemical carcinogens to induce transformation of cells derived from Syrian hamster embryo cultures can be determined quantitatively in vitro (1,2). Transformed colonies are characterized by a random criss-cross pattern of cells not seen in control colonies.…”

mentioning

confidence: 99%

X-Irradiation Enhancement of Transformation by Benzo(a)Pyrene in Hamster Embryo Cells

DrPaolo

Donovan

Nelson

1971

Proc. Natl. Acad. Sci. U.S.A.

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The addition of benzo(a)pyrene to cells obtained from Syrian hamster embryo cultures that had been plated to produce discrete colonies resulted in transformation. Altered colonies were not seen in controls. Prior irradiation of hamster embryo cells increased the transformation by benzo(a)pyrene, both per cell and per total colony. Maximum enhancement occurred when benzo(a)pyrene was added 48 hr after irradiation. Enhancement was greatest after 250-500 R, and decreased with higher doses of radiation. Although the link between radiation (both x-and UVirradiation) and cancer has been recognized since the discoveries of x-rays and natural radioactivity, the use of physical agents as carcinogens or cocarcinogens for in vitro studies has been extremely limited. X-irradiation has been shown to promote in vitro transformation in combination with polyoma (4) or simian virus 40 (5, 6) in hamster embryonic secondary cultures, hamster cell line BHK 21/13, and 3T3 mouse cell lines. Evidence was presented that the effect in established cell lines was not due to selection of preexisting cells with a greater susceptibility to transformation. Cell transformation in vitro by x-irradiation only has been reported in primary cell lines of hamster embryo cells (7), but has not been observed in the irradiated controls of the experiments involving radiation and viruses or after repeated weekly irradiation of a hypotetraploid fibroblast line derived from mouse spleen (8).Abbreviation: BP, benzo(a)pyrene. MATERIALS AND METHODSThe tissue-culture fluids and medium as well as the feeder layer cultures, the source and culture of cells from minced hamster embryos, and the standard conditions for the transformation assay have been described in detail (2). Cells derived from whole rat or hamster fetuses were grown as monolayers in Dulbecco's modification of Eagle's medium (MEM) with either 10% calf serum or 10% fetal bovine serum. In all experiments 2-to 4-day-old secondary or tertiary hamster cultures obtained by seeding 5 X 106 cells/100 ml plastic dish were used. Feeder layers composed of secondary cultures of rat embryo cells were irradiated as confluent monolayer cultures in 2 ml of medium. A Picker Portable Industrial x-ray apparatus (T55-433) was used at 110 kV and 5 mA, 2225 R/min, adjusted so that a total of 5000 R in air was delivered to a distance of 22.2 cm. Hamster cells were irradiated as confluent monolayers in 100-mm plastic Petri dishes with a Westinghouse Quandrocondex machine with two tubes, above and below, with the following factors: 200 kV (CP), 0.25 Cu + 0.55 mm Al filtration in each tube (HVL 0.9 mm Cu), 15 mA, 54 cm TSD, output 139 R/min. After being irradiated with 150-1000 R, the cells were plated on irradiated rat fibroblast feeder layers in 60-mm Petri dishes. Each value represents data obtained from at least 12 plates. Because unirradiated and irradiated cells had different survival rates, it was necessary to vary the number of hamster cells seeded to obtain the same approximate number of colonies. At desig...

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“…Although many mesenchymal-cell transformation systems are available (e.g. Chen and Heidelberger, 1969;DiPaolo et al, 1969DiPaolo et al, , 1972aRhim and Huebner, 1973;Evans and DiPaolo, 1975), epithelial models may provide a better understanding of the development and biological properties of the common adult human tumours, which are mostly of epithelial origin. To study neoplastic transformation in vitro, it is essential to identify changes in the in vitro properties of cells which correlate with tumorigenicity in vivo.…”

mentioning

confidence: 99%

Ultrastructure and biological markers of neoplastic change in adult mouse epithelial cells transformed in vitro

Knowles¹,

Franks²

1978

Br J Cancer

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Summary.-.The ultrastructure and in vitro growth properties of 5 tumorigenic mouse submandibular-gland epithelial cell lines were studied. In all lines, in vitro acinus formation occurred, and well differentiated epithelial cells showing epithelial microvilli and desmosomes and cytoplasmic tonofilaments were present. None of the cells showed specific ultrastructural features of the normal differentiated submandibular-gland ducts.All the lines formed colonies in semi-solid agar and on confluent monolayers of BALB/c 3T3 cells, and all lacked density-dependent inhibition of growth, as demonstrated by a random distribution of [3H]TdR labelling throughout growing colonies. These 3 growth properties appear to be reliable in vitro markers for epithelial neoplastic transformation in this system, but colony-forming efficiency in agar is lower than that reported for many transformed mesenchymal cells.

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Sarcoma-Producing Cell Lines Derived from Clones Transformed in vitro by Benzo[ a ]pyrene

Cited by 64 publications

References 4 publications

In vitro Malignant Transformation by Methylcholanthrene of the Progeny of Single Cells Derived from C3H Mouse Prostate

In vitro Malignant Transformation by Methylcholanthrene of the Progeny of Single Cells Derived from C3H Mouse Prostate

X-Irradiation Enhancement of Transformation by Benzo(a)Pyrene in Hamster Embryo Cells

Ultrastructure and biological markers of neoplastic change in adult mouse epithelial cells transformed in vitro

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