Excision-Repair in Primary Cultures of Mouse Embryo Cells and Its Decline in Progressive Passages and Established Cell Lines

Ben-Ishai, R.; Peleg, Leah

doi:10.1007/978-1-4684-2898-8_29

Cited by 21 publications

(15 citation statements)

References 8 publications

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“…We have found that the extraembryonic mesoderm represented the highest level of UDS yet found in any cultured rodent cell line or primary murine cells, whereas fetal skin had UDS that was about 10% of human levels, a level consistent with other studies [17,18]. The adult mouse skin cells represented the lowest level of UDS in this study (less than 1% of human UDS), which is consistent with other studies of mouse skin [17,18].…”

Section: Discussionsupporting

confidence: 91%

“…6B). This lineage had a mean of 9 grains/nucleus (37% of foreskin fibroblasts), representing a level of UDS that was 3-fold higher than the next highest level of UDS documented [17,18] namely that of the primary fetal skin fibroblasts (12.5% FF) (Fig. 6A).…”

Section: Resultsmentioning

confidence: 96%

“…Human foreskin fibroblast (ff) cells were included in all experiments to serve as a standard for comparison for all of the mouse cell lineages. Adult mouse skin was also included in these studies to serve as a reference point for a level of UDS that has been previously documented in murine skin fibroblasts [17,18,32,33].…”

Section: Resultsmentioning

confidence: 99%

“…The pattern or role of NER has never been defined in specific lineages during mammalian development, despite the fact that several human DNA repair deficiency syndromes manifest developmental abnormalities [16]. Studies of NER in mixed lineages during development using the functional assay for NER, UDS, have shown that in contrast to established mouse cell lines, primary murine embryonic fibroblasts excise dimers at a greater rate [17,18] and perform higher levels of repair synthesis [19]. Early passages of mixed embryonic fibroblasts have been shown to manifest higher levels of excision repair than later passages [17].…”

Section: Introductionmentioning

confidence: 99%

“…Studies of NER in mixed lineages during development using the functional assay for NER, UDS, have shown that in contrast to established mouse cell lines, primary murine embryonic fibroblasts excise dimers at a greater rate [17,18] and perform higher levels of repair synthesis [19]. Early passages of mixed embryonic fibroblasts have been shown to manifest higher levels of excision repair than later passages [17]. Peleg et al [18] have since shown that the capacity for excision repair of UV radiation damage in passaged embryonic fibroblasts from mouse is dependent upon the gestational stage and the duration of in vitro growth.…”

Section: Introductionmentioning

confidence: 99%

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Elevated DNA Excision Repair Capacity in the Extraembryonic Mesoderm of the Midgestation Mouse Embryo

Latimer

Hultner

Cleaver

et al. 1996

Experimental Cell Research

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In order to determine whether there is differential cell-type-specific DNA repair we measured the nucleotide excision repair capacity of the four distinct cell lineages that comprise the extraembryonic yolk sac using the unscheduled DNA synthesis assay. Yolk sacs from mouse embryos at 11.5-12.5 days gestation were microdissected to yield purified trophoblast, parietal endoderm, mesoderm, and visceral endoderm, as well as fetal skin fibroblasts which were then grown as primary explants. At this midgestational stage of development, the yolk sac provides essential functions for the sustenance of the embryo while the complex process of organogenesis is proceeding in the liver, kidney, and gut. Trophoblast giant cells, parietal endoderm, and visceral endoderm all demonstrated low levels of unscheduled DNA synthesis consistent with levels measured in adult mouse skin fibroblasts. As has previously been documented, embryonic mouse skin fibroblasts were reproducibly 2-to 3-fold higher than adult mouse skin fibroblasts in levels of DNA excision repair. The extraembryonic mesoderm, however, displayed a statistically significant level of unscheduled DNA synthesis 10-fold higher than adult mouse skin fibroblasts or the other lineages of the midgestation yolk sac. Further, the S-indexes of these lineages were also determined to assess the possible relevance of differential repair to the proliferative status of the cells. These data demonstrate that DNA excision repair capacity is lineage-specific during embryogenesis in the mouse. These studies may begin to provide a context for understanding the perplexing developmental aspects such as the characteristic congenital abnormalities associated with the human heritable DNA repair deficiency diseases.

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Section: Discussionsupporting

confidence: 91%

Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Elevated DNA Excision Repair Capacity in the Extraembryonic Mesoderm of the Midgestation Mouse Embryo

Latimer

Hultner

Cleaver

et al. 1996

Experimental Cell Research

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Revisiting the rodent repairadox

Hanawalt

2001

Environ and Mol Mutagen

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Cultured rodent and human cells typically display similar clonal survival characteristics following exposure to ultraviolet light (UV). However, compared to human cells, cultured cells from mice, rats, and hamsters are generally deficient in excision repair of the most prominent DNA lesion produced by UV, the cyclobutane pyrimidine dimer. In light of recent studies on the control of nucleotide excision repair, we are beginning to understand the basis for this so-called "repairadox." The resolution of this issue is important because rodents are so widely employed as surrogates for humans in genetic toxicology. This article will review the evolution in our understanding of rodent DNA repair and will also "revisit" my early association with my graduate mentor and esteemed colleague, Dick Setlow, in his honor upon the attainment of his 80th birthday.

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Metabolic activation of diesel exhaust carcinogens in primary and immortalized human TP53 knock‐in (Hupki) mouse embryo fibroblasts

Kucab

Phillips

Arlt

2012

Environ and Mol Mutagen

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Approximately 50% of human tumors have a mutation in TP53. The pattern and spectra of TP53 mutations often differ between cancer types, perhaps due to different etiological factors. The Hupki (human TP53 knock-in) mouse embryo fibroblast (HUF) immortalization assay is useful for studying mutagenesis in the human TP53 gene by environmental carcinogens. Prior to initiating an immortalization assay, carcinogen treatment conditions must be optimized, which can require a large number of cells. As primary HUF cultures senesce within 2 weeks, restricting their use, we investigated whether immortalized HUFs retaining wild-type TP53 can be surrogates for primary HUFs in initial treatment optimization. DNA damage by eight compounds found in diesel exhaust, benzo[a]pyrene, 3-nitrobenzanthrone, 1-nitropyrene, 1,3-dinitropyrene, 1,6-dinitropyrene, 1,8-dinitropyrene, 6-nitrochrysene, and 3-nitrofluorene, was assessed by (32) P-postlabeling and the alkaline comet assay in primary HUFs and in an immortal HUF cell line J201. For most compounds, higher levels of DNA adducts accumulated in J201 cells than in primary HUFs. This difference was not reflected in the comet assay or by cell viability changes. Experiments in three additional immortal HUF cell lines (AAI49, U56, and E2-143) confirmed strong differences in DNA adduct levels compared with primary HUFs. However, these did not correlate with the protein expression of Nqo1 or Nat1/2, or with gene expression of Cyp1a1 or Cyp1b1. Our results show that using immortal HUFs as surrogates for primary HUFs in genotoxicity screening has limitations and that DNA adduct formation is the best measure of genotoxicity of the nitro-polycyclic aromatic hydrocarbons tested in HUFs.

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Excision-Repair in Primary Cultures of Mouse Embryo Cells and Its Decline in Progressive Passages and Established Cell Lines

Cited by 21 publications

References 8 publications

Elevated DNA Excision Repair Capacity in the Extraembryonic Mesoderm of the Midgestation Mouse Embryo

Elevated DNA Excision Repair Capacity in the Extraembryonic Mesoderm of the Midgestation Mouse Embryo

Revisiting the rodent repairadox

Metabolic activation of diesel exhaust carcinogens in primary and immortalized human TP53 knock‐in (Hupki) mouse embryo fibroblasts

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