Labelling of DNA and differential sister chromatid staining after BrdU treatment in vivo

Pera, Franz; Mattias, P.

doi:10.1007/bf00292946

Cited by 53 publications

(5 citation statements)

References 7 publications

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“…Cell tracking has been a focus of active research in regenerative engineering. , Enhanced green fluorescent protein (EGFP) reporter gene, , luciferase reporter gene, − and 5-bromodeoxyuridine , are often used to image engineered cells. The luciferase and EGFP reporter genes, which have been used extensively in tumor cells, are rarely employed in implantology.…”

Section: Introductionmentioning

confidence: 99%

Light-Controlled BMSC Sheet–Implant Complexes with Improved Osteogenesis via an LRP5/β-Catenin/Runx2 Regulatory Loop

Jiang

Wang

et al. 2017

ACS Appl. Mater. Interfaces

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The combination of bone marrow mesenchymal stem cell (BMSC) sheets and titanium implants (BMSC sheet-implant complexes) can accelerate osseointegration. However, methods of fabricating BMSC sheet-implant complexes are quite limited, and the survival of BMSC sheet-implant complexes is one of the key barriers. Here, we show that a light-controlled fabricating system can generate less injured BMSC sheet-implant complexes with improved viability and osteogenesis and that noninvasive monitoring of the viability of BMSC sheet-implant complexes using a lentiviral delivery system is feasible. Enhanced green fluorescent protein- and luciferase-expressing BMSC sheets were used to track the viability of BMSC sheet-implant complexes in vivo. The experiments of micro-computed tomography analysis and hard tissue slices were performed to evaluate the osteogenic ability of BMSC sheet-implant complexes in vivo. The results showed that BMSC sheet-implant complexes survived for almost 1 month after implantation. Notably, BMSC sheet-implant complexes fabricated by the light-controlled fabricating system had upregulating expression levels of low-density lipoprotein-receptor-related protein 5 (LRP5), β-catenin, and runt-related transcription factor 2 (Runx2) compared to the complexes fabricated by mechanical scraping. Furthermore, we found that Runx2 directly bound to the rat LRP5 promoter and the LRP5/β-catenin/Runx2 regulatory loop contributed to the enhancement of the osseointegrating potentials. In this study, we successfully fabricated BMSC sheet-implant complexes with improved viability and osteogenesis and established a feasible, noninvasive, and continuous method for tracking BMSC sheet-implant complexes in vivo. Our findings lay the foundation for the application of BMSC sheet-implant complexes in vivo and open new avenues for engineered BMSC sheet-implant complexes.

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Section: Introductionmentioning

confidence: 99%

Light-Controlled BMSC Sheet–Implant Complexes with Improved Osteogenesis via an LRP5/β-Catenin/Runx2 Regulatory Loop

Jiang

Wang

et al. 2017

ACS Appl. Mater. Interfaces

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“…Differential staining of sister chromatids based on the incorporation of 5-bromodeoxyuridine (BrdUrd) into DNA [Goto et al, 1975;Ikushima and Wolff, 1974;Korenberg and Freedlender, 1974; Latt, 1973;Perry and Wolff, 1974;Scheres et al, 19771 has permitted the development of various systems for detecting and analyzing the effect of physical and chemical mutagens on the induction of sister chromatid exchanges (SCEs) in vivo [Allen and Latt, 1976;Allen et al, 1977;Bloom and Hsu, 1975;Comer et al, 1978;Kanda and Kato, 1979;Kligerman and Bloom, 1976;Morales-Ramirez, 1980;Pera and Mattias, 1976;Schneider et al, 1976;Vogel and Bauknecht, 19761, as well as in vitro [Guerrero et al, 1979;Latt, 1974;Perry and Evans, 1975;Raffetto et al, 1979;Stetka and Wolff, 19761. These systems are quite sensitive to chemical mutagens [Abe and Sasaki, 1977;Nakanishi and Schneider, 1979;Perry and Evans, 1975;Popescu et al, 19771 and ultraviolet (UV) light [MacRae et al, 1979;Popescu et al, 19791 but contradictory data have been obtained with ionizing radiation.…”

Section: Introductionmentioning

confidence: 99%

Effect of BrdUrd and low doses of gamma radiation on sister chromatid exchange, chromosome breaks, and mitotic delay in mouse bone marrow cells in vivo

1983

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We have measured, in mouse bone marrow cells in vivo, the ability of low doses of gamma radiation to induce sister chromatid exchanges (SCEs) in unifilarly 5-bromodeoxyuridine (BrdUrd)-substituted DNA. Also examined was the effect of BrdUrd incorporation on SCE induction by radiation, the comparative frequency of SCE and chromosome breaks induced by gamma radiation, the ability of ionizing radiation to interfere with normal cellular proliferation, and the relationship between proliferative inhibition and SCE and chromosome break frequency. A direct relationship between the number of SCEs and gamma radiation dose was observed, an effect which was dependent on BrdUrd incorporation. The frequency of SCE was 80 times higher than that of chromosome aberrations in cells with BrdUrd-substituted DNA, and there was no difference between the frequency of SCE in cells with or without chromosome breaks. The mitotic delay increased with the time between irradiation and harvesting. There was no relationship between the extent of mitotic delay and the number of SCEs.

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“…Two hours later, the cells were collected in order to accumulate metaphases. Cells were treated as described by Pera and Mattias (1976) to allow recognition of SCE between differentially stained chromatids. For each subject, 4 tubes were used and 30 intact metaphases were analyzed.…”

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confidence: 99%

Sister chromatid exchanges and chromosomal aberrations in lymphocytes of nurses handling cytostatic agents

Benhamou

Pot-Deprun

Sancho‐Garnier

et al. 1988

Intl Journal of Cancer

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A cohort study of 29 nurses who constantly handled cytostatic drugs, and 29 controls matched according to sex and age, was carried out between 1983 and 1986. Cytogenetic damage was assessed by sister chromatid exchanges (SCE) and chromosomal aberrations. No significant increase in mean number of SCE was found for nurses (7.37) as compared to matched controls (7.00), whereas a significant excess of SCE (p less than 0.001) was observed for smokers (8.23) as compared to non-smokers (6.75). The number of SCE was studied in relation to the amount and nature of cytostatics handled as well as to the duration of exposure. A significant association (p less than 0.05) was found between individual mean number of SCE and the total number of drugs handled after adjustment for confounding factors. In contrast, the number of SCE was not significantly related to the nature of drugs handled or to the duration of exposure. With regard to chromosomal damage, no significant difference was observed between nurses and controls in gap, break, dicentric and translocation frequencies.

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Labelling of DNA and differential sister chromatid staining after BrdU treatment in vivo

Cited by 53 publications

References 7 publications

Light-Controlled BMSC Sheet–Implant Complexes with Improved Osteogenesis via an LRP5/β-Catenin/Runx2 Regulatory Loop

Light-Controlled BMSC Sheet–Implant Complexes with Improved Osteogenesis via an LRP5/β-Catenin/Runx2 Regulatory Loop

Effect of BrdUrd and low doses of gamma radiation on sister chromatid exchange, chromosome breaks, and mitotic delay in mouse bone marrow cells in vivo

Sister chromatid exchanges and chromosomal aberrations in lymphocytes of nurses handling cytostatic agents

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