Combined introduction of Bmi-1 and hTERT immortalizes human adipose tissue-derived stromal cells with low risk of transformation

Tátrai, Péter; Szepesi, Áron; Matula, Zsolt; Szigeti, Anna; Buchan, Gyöngyi; Mádi, András; Uher, Ferenc; Német, Katalin

doi:10.1016/j.bbrc.2012.04.088

Cited by 47 publications

(49 citation statements)

References 22 publications

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“…Lentiviral particles were produced in HEK293T cells transfected by the calcium phosphate co-precipitation method according to manufacturer’s protocol [25]. Viral titers of the supernatants were estimated by flow cytometric measurement of HEK cells infected with a dilution series of the supernatants; transduced cells were selected with 2 ug/ml puromycin for 2 weeks.…”

Section: Methodsmentioning

confidence: 99%

Screening the Expression of ABCB6 in Erythrocytes Reveals an Unexpectedly High Frequency of Lan Mutations in Healthy Individuals

et al. 2014

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Lan is a high-incidence blood group antigen expressed in more than 99.9% of the population. Identification of the human ABC transporter ABCB6 as the molecular basis of Lan has opened the way for studies assessing the relation of ABCB6 function and expression to health and disease. To date, 34 ABCB6 sequence variants have been described in association with reduced ABCB6 expression based on the genotyping of stored blood showing weak or no reactivity with anti-Lan antibodies. In the present study we examined the red blood cell (RBC) surface expression of ABCB6 by quantitative flow cytometry in a cohort of 47 healthy individuals. Sequencing of the entire coding region of the ABCB6 gene in low RBC ABCB6 expressors identified a new allele (IVS9+1G>A, affecting a putative splice site at the boundary of exon 9) and two nonsynonymous SNPs listed in the SNP database (R192Q (rs150221689) and G588 S (rs145526996)). The R192Q mutation showed co-segregation with reduced RBC ABCB6 expression in a family, and we found the G588 S mutation in a compound heterozygous individual with undetectable ABCB6 expression, suggesting that both mutations result in weak or no expression of ABCB6 on RBCs. Analysis of the intracellular expression pattern in HeLa cells by confocal microscopy indicated that these mutations do not compromise overall expression or the endolysosomal localization of ABCB6. Genotyping of two large cohorts, containing 235 and 1039 unrelated volunteers, confirmed the high allele frequency of Lan-mutations. Our results suggest that genetic variants linked to lower or absent cell surface expression of ABCB6/Langereis may be more common than previously thought.

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Section: Methodsmentioning

confidence: 99%

Screening the Expression of ABCB6 in Erythrocytes Reveals an Unexpectedly High Frequency of Lan Mutations in Healthy Individuals

et al. 2014

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“…Total RNA was isolated from undifferentiated and differentiated cells and cDNA was synthesized as previously described [20]. The expression levels of OCT4, SOX2, NANOG, ALP, ABCG2, CEMP1, and the peroxisome-proliferatoractivated receptor gamma (PPARg) gene known as an adipocyte marker were measured using TaqMan reagents.…”

Section: Gene Expression Analysismentioning

confidence: 99%

“…Expression of the cell surface markers was assessed by flow cytometry, as described previously [20]. All antibodies were purchased from BD Biosciences (Erembodegem, Belgium).…”

Section: Flow Cytometrymentioning

confidence: 99%

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ABCG2 Is a Selectable Marker for Enhanced Multilineage Differentiation Potential in Periodontal Ligament Stem Cells

Szepesi

Matula

Szigeti

et al. 2015

Stem Cells and Development

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“…Telomerase is expressed in highly proliferating cells throughout the developing embryo, while it is dramatically down-regulated as cells differentiate, being undetectable in many adult somatic cells242526. Otherwise, studies on cancer cells and engineered somatic cells overexpressing telomerase have shown that this enzyme can confer an immortal phenotype2728. Therefore, developing a tool that can revert the molecular mechanisms underlying stem cell senescence in vitro may pave the way to affordable strategies of stem cell expansion, improving the outcome of cell therapy efforts.…”

mentioning

confidence: 99%

Stem cell senescence. Effects of REAC technology on telomerase-independent and telomerase-dependent pathways

Rinaldi

Maioli

Pigliaru

et al. 2014

Sci Rep

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Decline in the gene expression of senescence repressor Bmi1, and telomerase, together with telomere shortening, underlay senescence of stem cells cultured for multiple passages. Here, we investigated whether the impairment of senescence preventing mechanisms can be efficiently counteracted by exposure of human adipose-derived stem cells to radio electric asymmetrically conveyed fields by an innovative technology, named Radio Electric Asymmetric Conveyer (REAC). Due to REAC exposure, the number of stem cells positively stained for senescence associated β-galactosidase was significantly reduced along multiple culturing passages. After a 90-day culture, REAC-treated cells exhibited significantly higher transcription of Bmi1 and enhanced expression of other stem cell pluripotency genes and related proteins, compared to unexposed cells. Transcription of the catalytic telomerase subunit (TERT) was also increased in REAC-treated cells at all passages. Moreover, while telomere shortening occurred at early passages in both REAC-treated and untreated cells, a significant rescue of telomere length could be observed at late passages only in REAC-exposed cells. Thus, REAC-asymmetrically conveyed radio electric fields acted on a gene and protein expression program of both telomerase-independent and telomerase-dependent patterning to optimize stem cell ability to cope with senescence progression.

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Combined introduction of Bmi-1 and hTERT immortalizes human adipose tissue-derived stromal cells with low risk of transformation

Cited by 47 publications

References 22 publications

Screening the Expression of ABCB6 in Erythrocytes Reveals an Unexpectedly High Frequency of Lan Mutations in Healthy Individuals

Screening the Expression of ABCB6 in Erythrocytes Reveals an Unexpectedly High Frequency of Lan Mutations in Healthy Individuals

ABCG2 Is a Selectable Marker for Enhanced Multilineage Differentiation Potential in Periodontal Ligament Stem Cells

Stem cell senescence. Effects of REAC technology on telomerase-independent and telomerase-dependent pathways

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