Karyotypic stability of Serum-Free Mouse Embryo (SFME) cells

Ernst, Ted; Jackson, Constance; Barnes, David W.

doi:10.1007/bf00556291

Cited by 11 publications

(5 citation statements)

References 19 publications

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“…These serum-free mouse embryo (SFME) 1 cells exhibit a number of unusual properties. Unlike mouse embryo cells derived in conventional, serum-supplemented media which undergo growth crisis and immortalization, SFME cells do not lose proliferative potential or develop gross chromosomal aberration when cultured for more than 10 times the number of population doublings that can be achieved with mouse embryo cells in conventional, serum-containing medium (Todaro and Green, 1963;Loo et al, 1987Loo et al, , 1989bErnst et al, 1990).…”

mentioning

confidence: 99%

Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation.

Rawson

Loo

Duimstra

et al. 1991

The Journal of Cell Biology

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Abstract. Serum-free mouse embryo (SFME) cells, derived in medium in which serum is replaced with growth factors and other supplements, are proastroblasts that are acutely dependent on epidermal growth factor (EGF) for survival. Ultrastructurally, an early change found in SFME cells deprived of EGF was a loss of polysomes which sedimentation analysis confirmed to be a shift from polysomes to monosomes. The ribosomal shift was not accompanied by decreased steady-state level of cytoplasmic actin mRNA examined as an indicator of cellular mRNA level. With time the cells became small and severely degenerate and exhibited nuclear morphology characteristic of apoptosis. Genomic DNA isolated from cultures undergoing EGF deprivation-dependent cell death exhibited a pattern of fragmentation resulting from endonuclease activation characteristic of cells undergoing apoptosis or programmed cell death. Flow cytometric analysis indicated that cultures in the absence of EGF contained almost exclusively Gl-phase cells. Some of the phenomena associated with EGF deprivation of SFME cells are similar to those observed upon NGF deprivation of nerve cells in culture, suggesting that these neuroectodermal-derived cell types share common mechanisms of proliferative control involving peptide growth factor-dependent survival.

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confidence: 99%

Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation.

Rawson

Loo

Duimstra

et al. 1991

The Journal of Cell Biology

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“…SFME cells were derived and are maintained in serum-free medium supplemented with EGF and other growth factors. Although they have been passaged in culture for years, SFME cells have not undergone a detectable growth crisis and exhibit karyotypic stability (15). The presence of serum is growth restrictive and results in G 1 arrest.…”

Section: Discussionmentioning

confidence: 99%

Simian Virus 40 Large T Antigen J Domain and Rb-Binding Motif Are Sufficient To Block Apoptosis Induced by Growth Factor Withdrawal in a Neural Stem Cell Line

Slinskey

Barnes

Pipas

1999

J Virol

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Serum-free mouse embryo (SFME) cells are a neural stem cell line that is dependent upon epidermal growth factor (EGF) for survival. Removal of EGF results in the G1 arrest and apoptosis of SFME cells. We have shown that the expression of simian virus 40 large T antigen in SFME cells blocks apoptosis and allows cell survival and division in the absence of EGF. Therefore the presence of T antigen abrogates the EGF requirement. The steady-state levels of p53, p21, and mdm-2 do not increase as SFME cells undergo apoptosis upon EGF withdrawal. Furthermore, the amino-terminal 136 amino acids (N136) of T antigen are sufficient to block death and to promote proliferation in the absence of EGF, while the carboxy-terminal fragment (C251–708), which contains the p53 binding site, is unable to block death. Taken together, these data suggest that SFME cells deprived of EGF undergo p53-independent apoptosis. Mutations that disrupt either the J domain or Rb family binding abolish the ability of T antigen to block SFME cell apoptosis and to promote cell growth. We conclude that T antigen must act on one or more members of the Rb family to inhibit SFME cell apoptosis.

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“…It is unclear at present why astrocytes differentiated from SFME cells lacks the mechanisms of growth inhibition by TGFβ. On the other hand, SFME cells were maintained in serum-free medium supplemented with EGF and other growth factors (Ernst et al, 1991). SFME cells arrested growth at the G1 phase of cell cycle and subsequently underwent apoptosis upon the withdrawal of EGF (Rawson et al, 1990).…”

Section: Tgf-β Induced Differentiation Of Neural Stem Cells Into Astrocytesmentioning

confidence: 99%

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et al. 2001

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Serum-free mouse embryo (SFME) cells were established by D. Barnes et al., and are known to be a neural stem cell line, which differentiate into astrocytes upon treatment with TGF-beta. Therefore, SFME cells is thought to be a model well suited to analyze the differentiation mechanism of neural stem cells. Until now, we have investigated the regulation mechanisms of telomerase activity and telomere length in human cancer and normal cells. Telomerase is the enzyme responsible for the synthesis and maintenance of telomere repeats located at chromosomal ends and is normally expressed in embryonic and germline cells, but not in most normal cells. Here, using SFME cells, we attempted to analyze the regulation mechanism of telomerase activity in neural stem cells and to detect a change upon differentiation into astrocytes. When SFME cells were cultured in the presence of TGF-beta, cells showed anelongated morphology and decreased its growth to 50% of control culture. Cells also expressed the glial fibrillary acidic protein (GFAP), a marker for astrocytes,indicating that TGF-beta induced differentiation in SFME cells from neural stem cells into astrocytes. At the same time,TGF-beta also inhibited telomerase activity and repressed the expression of the mouse telomerase reverse transcriptase(mTERT), demonstrating that SFME cells was vested with a finite replicative life span upon treatment with TGF-beta. To understand the mechanisms regulating mTERT levels during differentiation into astrocytes, we have estimated the expression level of c-myc, which is known to be a key molecule in activating the TERT promoter. As a result, TGF-beta-treated SFME cells were shown to repress the expression of c-myc. Furthermore, promoter analysis, using the 5'-region of the mTERT gene, which possess two E-box elements bound to c-Myc/Max, demonstrated that mTERT promoter activity greatly decreased in TGF-beta-treated SFME cells as compared to non-treated SFME cells. These suggest that c-myc might play a critical role in the expression of mTERT, and that down-regulation of c-myc dependent upon the astrocytic differentiation in SFME cells might cause the repression of mTERT in TGF-beta-treated SFME cells.

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Karyotypic stability of Serum-Free Mouse Embryo (SFME) cells

Cited by 11 publications

References 19 publications

Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation.

Death of serum-free mouse embryo cells caused by epidermal growth factor deprivation.

Simian Virus 40 Large T Antigen J Domain and Rb-Binding Motif Are Sufficient To Block Apoptosis Induced by Growth Factor Withdrawal in a Neural Stem Cell Line

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