Human brain in tissue culture. III. PML‐SV40‐induced transformation of brain cells and establishment of permanent lines

Santoli, Daniela; Wróblewska, Zofia; Gilden, Donald H.; Girardi, Anthony J.; Koprowski, Hilary

doi:10.1002/cne.901610304

Cited by 23 publications

(3 citation statements)

References 19 publications

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“…JCV genomic DNA was prepared from the plasmid pJC (15) purified after BamHI and Hha I digestion, and passage over a Sephacryl S-1000 column (Pharmacia). The pSV2 vector was constructed by Mulligan and Berg (16) gin-defective mutant of SV40 (mutant [1][2][3][4][5][6][7][8][9][10][11] was provided by Y. Gluzman as an insert in the pMK16 vector at the BamHI site. All plasmids were grown in Escherichia coli K-12 hosts, amplified using chloramphenicol, and purified by a cleared lysis method (17,18).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, Miranda et al (8) have demonstrated that homogenous populations of human myoblast cells could be isolated from skeletal muscle cells after transformation with simian tumor virus 40 (SV40). Santoli et al (9) and Shein (10) have transformed human adult brain and fetal glial cells, respectively, using SV40. However, infectious SV40 was shed from these cells and, in the case of the fetal brain, the life span of the cells lasted only 13-39 cell passages.…”

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confidence: 99%

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Establishment of a line of human fetal glial cells that supports JC virus multiplication.

Major¹,

Miller²,

Mourrain³

et al. 1985

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

305

247

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Primary cultures of human fetal brain cells were transfected with plasmid DNA pMK16, containing an origin-defective mutant of simian virus 40 (SV40). Several weeks after DNA treatment, proliferation of glial cells was evident in the culture, allowing passage of the cells at low split ratios. Initially, only 10% of the cells demonstrated nuclear fluorescence staining using a hamster tumor antibody to the SV40 T protein. By the sixth passage, however, 100% of the cells reacted positively to the same antibody. During these early passages, the cells designated SVG began growing very rapidly and acquired a homogenous morphology. Cell division required only low serum concentrations, was not contact-inhibited, and remained anchorage dependent. These characteristics of the SVG cells have been stable through 25 passages or -80 cell generations. The SV40 T protein is continuously produced in the cells and can direct the replication of DNA inserts in the pSV2 vector, determined by in situ hybridization using biotinlabeled DNA probes, which contains the SV40 replication origin. More importantly, SVG cells support the multiplication of the human papovavirus JCV at levels comparable to primary cultures of human fetal glial cells, producing infectious virus as early as 1 week after viral adsorption. Their brain-cell derivation has been established as astroglial, based on their reactivity with a monoclonal antibody to glial fibrillary acid protein and lack of activity with an anti-galactocerebroside antibody, which identifies oligodendroglial cells. The SVG cells represent a unique line of continuous rapidly growing human fetal astroglial cells that synthesizes a replication-proficient SV40 T protein. Their susceptibility to JC virus (JCV) infection obviates a host restriction barrier that limited JCV studies to primary cultures of human fetal brain and thus should allow for more detailed molecular studies of human brain cells and JCV that infects them.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Establishment of a line of human fetal glial cells that supports JC virus multiplication.

Major¹,

Miller²,

Mourrain³

et al. 1985

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

305

247

View full text Add to dashboard Cite

show abstract

“…There have been reports that certain cells are more susceptible to SV40-induced transformation. In early studies it was claimed th at the likelihood of obtaining permanent lines increased with the age of the diploid culture (Jensen et al 1963), but this could not be confirmed by other investigators (Santoli et al 1975;Huschtscha & Holliday 1983). In comparison with normal cells, a consistent increase in the number of SV40-induced foci was reported in skin fibroblasts from patients with Fanconi's anaemia, which is a disease with a high incidence of chromosome abnormalities and spontaneous neoplasms (Todaro 1966 ;Todaro 1968).…”

Section: Introductionmentioning

confidence: 94%

The susceptibility of Werner’s syndrome and other human skin fibroblasts to SV40-induced transformation and immortalization

Huschtscha¹,

Thompson²,

Holliday³

1986

Proc. R. Soc. Lond. B.

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In attempts to transform and immortalize human cell cultures, skin fibroblasts from normal donors of different ages, from patients with the premature ageing diseases Werner’s syndrome (WS) and progeria (PR), and from donors with the cancer-prone diseases ataxia telangiectasia (AT), Bloom’s syndrome (BS) and Fanconi’s anaemia (FA), were infected with SV40 virus and their growth monitored thereafter. Lesch–Nyhan (LN) fibroblasts were also infected. SV40-infected cultures from two normal and from WS, AT and LN donors attained a spectrum of transformed properties, high mitotic activity at confluence, presence of T-antigen, anchorage independence and altered morphology. Most of these pretransformed cultures died in the crisis period. However, two cultures from the WS and LN patients survived the crisis period and have now been grown to more than 200 passages. For the LN culture the crisis period was at least 200 days. Both permanent lines retain the properties of pretransformed cells, but differ in their modal chromosome number and ability to grow in methionine-free medium. It can be concluded from these experiments that transformation by SV40 to permanent lines is a rare event in human skin fibroblasts, even when these cells were taken from patients predisposed to form cancers.

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Human brain in tissue culture. IV. Morphological characteristics

Rorke

Gilden

Wróblewska

et al. 1975

J of Comparative Neurology

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Human brain in tissue culture. III. PML‐SV40‐induced transformation of brain cells and establishment of permanent lines

Cited by 23 publications

References 19 publications

Establishment of a line of human fetal glial cells that supports JC virus multiplication.

Establishment of a line of human fetal glial cells that supports JC virus multiplication.

The susceptibility of Werner’s syndrome and other human skin fibroblasts to SV40-induced transformation and immortalization

Human brain in tissue culture. IV. Morphological characteristics

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