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Journal ArticleDOI

Analysis of three whole-arm translocations in a mouse sarcoma cell line.

01 Jan 1984-Cytogenetic and Genome Research (Karger Publishers)-Vol. 38, Iss: 3, pp 161-164
TL;DR: The possibility that these whole arm translocations arose by telomeric fusion and the molecular basis of such fusions are discussed.
Abstract: Three apparently whole-arm translocation chromosomes in the mouse sarcoma 180 ascites cell line have been studied by G- and C-banding and by Hoechst staining. All three chromosomes appear to have two centromeres. Both centromeres of one, which are very close together, are likely to be active and to produce parallel separation of the chromatids. The centromeres of the other two chromosomes are well separated. One of these two centromeres may be inactive either because the kinetochore organizer has been inactivated or because the kinetochore plate has been deleted, leaving the AT-rich centromeric constituative heterochromatin intact. The possibility that these whole arm translocations arose by telomeric fusion and the molecular basis of such fusions are discussed.
Citations
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Journal ArticleDOI
TL;DR: Karyotype and DNA content of the cell line reveal a mostly euploid and rather stable cell population, which is of great interest for cytogenetic studies.

123 citations


Cites background from "Analysis of three whole-arm translo..."

  • ...In general, the adaptation of cell lines to in vitro conditions is often associated with dynamic chromosomal changes affecting chromosome number and/or morphology, giving rise in some cases to marker chromosomes which are line-specific (Ghosh and Chaudhuri, 1984)....

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Journal ArticleDOI
TL;DR: Long-term cultures of certain rat and mouse cell lines carry several dicentric and some multicentric chromosomes and the number of kinetochores formed along the length of these chromosomes is studied.
Abstract: Long-term cultures of certain rat and mouse cell lines carry several dicentric and some multicentric chromosomes. Using antikinetochore antibodies obtainable from serum of scleroderma (var. CREST) patients we studied the number of kinetochores formed along the length of these chromosomes. The rat cells displayed as many kinetochores as there were centromeres. However, mouse cells showed the synthesis of only one kinetochore in dicentric and multicentric chromosomes which had been in the culture for a period of 1 year or more. When translocations were induced by bleomycin in mouse L cells, the newly formed dicentric chromosomes showed the formation of two kinetochores. It is not known when the accessory centromeres lose their capacity to assemble kinetochore proteins. Possibly, in the rat the ‘latent’ kinetochores lack a specific component which renders them ineffective for microtubule binding. The reason for the formation of only one kinetochore in mouse multicentric chromosomes is not clear. It may be due to the accumulation of mutations, modification of the kinetochore protein so that it lacks the antibody binding component, or a more effective regulatory gene than in the rat.

28 citations

Journal ArticleDOI
TL;DR: The results indicate that telomersase has been activated in the course of SAF-1 development, and the highest levels of telomerase activity correlate with an increase in cell proliferation, thus supporting a permanent cell line.
Abstract: Fish cell lines are increasingly important research tools. The SAF-1 cell line, fibroblast-like culture derived from the marine fish gilthead seabream (Sparus aurata), has proved useful in many applications, especially in viral research. For cell lines intended as in vitro models, characterization of their properties and authentication are essential for deeper understanding of their performance and thus more precise experimental design and applicability. In this study we characterized the SAF-1 cell line in terms of genetic stability through time and genetic labeling. Methods for determining stability include telomerase activity, karyotyping, mapping of ribosomal RNA regions, and DNA content. For genetic labeling 12 microsatellite loci were used. The results indicate that telomerase has been activated in the course of SAF-1 development, and the highest levels of telomerase activity correlate with an increase in cell proliferation, thus supporting a permanent cell line. This stability is in agreement with the normal situation presented by the cytogenetic traits and DNA content values, and the genotypic profile allows SAF-1 authentication at the single individual level. This study increases the value of SAF-1 as an in vitro system, which is now one of the few well-characterized cell lines from a marine fish.

20 citations


Cites background from "Analysis of three whole-arm translo..."

  • ...Changes in the karyotype affecting chromosome number or morphology have often been associated with adaptation of cells to in vitro conditions (Ghosh and Chaudhuri, 1984)....

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  • ...The matching probability (Mp), as the number of individuals that may be surveyed before finding the same genotype in a randomly selected sample [15], has been calculated....

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Journal ArticleDOI
TL;DR: The utility of this cell line as an in vitro model for aquatic toxicological studies and the cell line will be further useful for studying oxidative stress markers against aquatic pollutants is illustrated.
Abstract: A new epithelial cell line, WAF was developed from caudal fin of freshwater shark, Wallago attu. The cell line was optimally maintained at 28 °C in Leibovitz-15 (L-15) medium supplemented with 20 % fetal bovine serum. The cell line was characterized by various cytogenetic and molecular markers. The cytogenetic analysis revealed a diploid count of 86 chromosomes at different passages. The origin of the cell lines was confirmed by the amplification of 547 and 654 bp sequences of 16S rRNA and cytochrome oxidase subunit I genes of mitochondrial DNA, respectively. WAF cells were characterized for their growth characteristics at different temperature and serum concentration. Epithelial morphology of the cell line was confirmed using immunocytochemistry. Further cell plating efficiency, transfection efficiency and viability of cryopreserved WAF cells was also determined. Cytotoxicity and genotoxicity assessment of cadmium salts on WAF cells by MTT, NR and comet assay illustrated the utility of this cell line as an in vitro model for aquatic toxicological studies. The cell line will be further useful for studying oxidative stress markers against aquatic pollutants.

10 citations

Dissertation
18 Jun 2017
TL;DR: Preliminary applications such as gene transfection studies and cytotoxicity assays using bacterial extracellular products were done in addition to ensuring that the cultures and cryostocks were contamination-free.
Abstract: Cell lines and primary cell cultures from fish tissues have been used for research in disease diagnosis and cytotoxicity evaluation of environmental pollutants. The principal aim of this work was the development of continuous cell lines from the caerulean damsel, Pomacentus caeruleus. To achieve this, tissues were first sourced from donor fishes, devoid of contaminants using standardised disinfection protocols. Primary cultures were then initiated and suitable media, additives, dissociation reagents as well as optimum incubation temperature were determined. Successful primary cultures were subcultured and passaged to derive continuous cell lines which were cryopreserved for long term storage. The continuous cell lines established were characterised by immunotyping using cell type markers and authenticated to confirm the species of origin using previously established barcoding techniques. Preliminary applications such as gene transfection studies and cytotoxicity assays using bacterial extracellular products were done in addition to ensuring that the cultures and cryostocks were contamination-free.

5 citations


Cites background from "Analysis of three whole-arm translo..."

  • ...In general, the adaptation of cell lines to in vitro conditions is often associated with dynamic chromosomal changes affecting chromosome number and/or morphology, giving rise in some cases to marker chromosomes which are line-specific (Ghosh and Chaudhuri, 1984)....

    [...]