The biology of human cells in tissue culture. I. Characterization of cells derived from osteogenic sarcomas

Smith, Helene S.; Owens, Robert B.; Hiller, Alan J.; Nelson‐Rees, Walter A.; Johnston, James O.

doi:10.1002/ijc.2910170211

Cited by 66 publications

(15 citation statements)

References 23 publications

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“…The modal number is 46, with two X chromosomes (39). When the karyotype of SV/HF-5 was investigated, it was found to represent a bimodal cell population with a group of pseudodiploid cells (greater than 50% of the population) having a chromosome range of 42 to 46 (modal number, 45) and a second group of heteroploid (pseudotetraploid) cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

Immortalization of human fibroblasts transformed by origin-defective simian virus 40.

Neufeld¹,

Ripley²,

Henderson³

et al. 1987

Mol. Cell. Biol.

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Simian virus 40 (SV40)-mediated transformation of human diploid fibroblasts has provided an effective experimental system for studies of both "senescence" in cell culture and carcinogenesis. Previous interpretations may have been complicated, however, by the semipermissive virus-cell interaction. In earlier studies, we previously demonstrated that the human diploid fibroblast line HS74 can be efficiently transformed by DNA from replication-defective mutants of SV40 containing a deletion in the viral origin for DNA synthesis (SVori-). In the current study, we found that such SVori-transformants show a significantly increased life span in culture, as compared with either HS74 or an independent transformant containing an intact viral genome, but they nonetheless undergo senescence. We have clonally isolated six immortalized derivatives of one such transformant (SV/HF-5). Growth studies indicate that the immortalized cell lines do not invariably grow better than SVWHF-5 or HS74. Genetic studies involving karyotypic analysis and Southern analysis of integrated viral sequences demonstrated both random and nonrandom alterations. All immortalized derivatives conserved one of the two copies of SV40 sequences which expressed a truncated T antigen. These cloned SV40-transformed cell lines, pre-and postimmortalization, should be useful in defining molecular changes associated with immortalization.

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Section: Resultsmentioning

confidence: 99%

Immortalization of human fibroblasts transformed by origin-defective simian virus 40.

Neufeld¹,

Ripley²,

Henderson³

et al. 1987

Mol. Cell. Biol.

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“…Indeed, efficient in vitro culture of normal human keratinocytes requires the presence of irradiated fibroblasts (29). Recent work also suggests that fibroblast abnormality detectable in vitro may be a concomitant of several different tumors (11).…”

Section: Methodsmentioning

confidence: 99%

Defective organization of actin in cultured skin fibroblasts from patients with inherited adenocarcinoma

Kopelovich

Conlon

Pollack

1977

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

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“…Primary human fibroblasts (HS74BM cells) (14) were used to test the transforming potential of the recombinant viruses. Figure 1 shows the morphology of the transformed foci.…”

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confidence: 99%

Efficient transformation of human fibroblasts by adenovirus-simian virus 40 recombinants.

Doren¹,

Gluzman²

1984

Mol. Cell. Biol.

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The origin-defective simian virus 40 (SV40) mutant 6-1 has been useful in transforming human cells (Small et al., Nature [London] 296:671-672, 1982; Nagata et al., Nature [London] 306:597-599, 1983). However, the low efficiency of transformation achieved by DNA transfection is a major drawback of the system. To increase the efficiency of SV40-induced transformation of human fibroblasts, we used recombinant adenovirus-SV40 virions which contain a complete SV40 early region including either a wild-type or defective (6-1) origin of replication. The SV40 DNA was cloned into the adenovirus vector in place of early region 1. Cell lines transformed by viruses containing a functional origin of replication produced free SV40 DNA. These cell lines were subcloned, and some of the subclones lost the ability to produce free viral DNA. Subclones that failed to produce free viral DNA were found to possess a mutated T antigen. Cell lines transformed by viruses containing origin-defective SV40 mutants did not produce any free DNA. Because of the high efficiency of transformation, we suggest that the origin-defective chimeric virus is a convenient system for establishing SV40-transformed cell lines from any human cell type that is susceptible to infection by adenovirus type 5.Cells range from fully permissive to fully nonpermissive for simian virus 40 (SV40) infection. Permissivity results in full expression of the viral genome, replication of viral DNA, and production of infectious virions. Nonpermissive infection results in expression of the early viral genes, but no viral DNA is synthesized or virions produced. Nonpermissive cells can be transformed, and the viral genome can be integrated into the host cell genome. Human cells are semipermissive for SV40. Virus production occurs to ca. 1% the level produced by permissive monkey kidney cells (12). The majority of viral DNA replication occurs in ca. 1 to 2% of the cells, where it reaches levels equivalent to wild-type infection (18). A portion of the population can become transformed and integrate viral DNA into the host cell genome (18); however, the efficiency is quite low. The frequency of transformation can be enhanced by transfection of SV40 origin-defective mutants (13). Because DNA transfection is an inefficient process, the frequency of transformation is still not very high. We wanted to determine whether we could increase the efficiency of SV40-induced tranformation after infection with recombinant viruses. We were also interested in determining whether having a functional versus a defective SV40 origin of replication would affect the frequency of transformation by recombinant viruses.The chimeric viruses contain the SV40 early region which encodes T antigen (HpaII-BamHI fragment) (16) cloned into the helper independent adenovirus vector, AE1/X (17). The SV40 origin of replication is either wild type or the orimutant 6-1, having six nucleotides deleted at the BglI site (4, 5). The SV40 DNA is cloned into the adenovirus vector in place of early regions la and lb. ...

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The biology of human cells in tissue culture. I. Characterization of cells derived from osteogenic sarcomas

Cited by 66 publications

References 23 publications

Immortalization of human fibroblasts transformed by origin-defective simian virus 40.

Immortalization of human fibroblasts transformed by origin-defective simian virus 40.

Defective organization of actin in cultured skin fibroblasts from patients with inherited adenocarcinoma

Efficient transformation of human fibroblasts by adenovirus-simian virus 40 recombinants.

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