studies with bromodeoxyuridine on L-strain mouse cells.

by Sop-Chong Kim in [Toronto]

Written in English
Published: Pages: 34 Downloads: 152
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Subjects:

  • Cells,
  • Bromodeoxyuridine,
  • Radio-biology
  • Edition Notes

    ContributionsToronto, Ont. University. Theses (M.Sc.)
    Classifications
    LC ClassificationsLE3 T525 MSC 1969 K56
    The Physical Object
    Pagination[34, 7 leaves]
    Number of Pages34
    ID Numbers
    Open LibraryOL18170938M

Quantification of the percentage of BrdU + α-cells (Fig. 5B) and δ-cells (Fig. 5D) showed STZ treatment significantly increased the proliferation of these non-β cells, supporting that proliferation was involved in the regeneration of these cells. It should be noted that a significant number of α-cells and δ-cells, including those that.   The BrdU labelling technique is primarily used in short-term endpoint assays, but in mammalian cells it can also be used over several cell cycles, and even in vivo, with BrdU being added to the drinking water of mice to monitor the turnover of cellular compartments such as haematopoietic and intestinal cells over weeks or months. BrdU is thus. RESULTS Loss of insulin-positive cells from cultures of mouse islets. To ask whether insulin-negative cells seen in islet cultures originate from differentiated β-cells, we used a genetic lineage tracing approach (), which allows identification of differentiated β-cells and their progeny via Cre-mediated recombination in transgenic first adapted the protocols established for adherent. Low concentrations (1–3 µg/ml) of 5-bromodeoxyuridine (BrdU) reversibly suppress pigmentation in a highly pigmented clone (B 5 59) of cultured B16 mouse melanoma cells. We have found that unpigmented cells (clone C 3 ), derived by long-term culture of B 5 59 cells in 1 µg of BrdU/ml, were completely amelanotic with no biochemically or cytochemically detectable tyrosinase activity or.

  The quantification of BrdU-positive cells was performed on digital images captured under × magnification of each thyroid section. Two randomly selected slides from three thyroids of each group were analyzed. For each slide, ratios of BrdU-positive cells/total cells from 10–12 bright fields under × amplification were calculated. JAX ® Mice are named according to the guidelines set by the International Committee on Standardized Genetic Nomenclature for Mice, and strain names are revised as necessary to conform to these guidelines.. Learn the highlights of mouse strain nomenclature with our short, interactive tutorial. Understanding appropriate nomenclature is essential due to the complexity of strain names for. Alveolar epithelial type II cells, a major source of angiotensin-converting enzyme (ACE)-2 in the adult lung, are normally quiescent but actively proliferate in lung fibrosis and downregulate this protective enzyme. It was, therefore, hypothesised that ACE-2 expression might be related to cell cycle progression. To test this hypothesis, ACE-2 mRNA levels, protein levels and enzymatic activity. Labeling Cells with Anti-BrdU and Propidium Iodide (PI) for Flow Cytometric or Microscopic Analysis Mouse or human cell suspension (see the Becton Dickinson procedures for preparation of cell suspensions, Monoclonal Antibodies Source Book, Sections and ) or tissue culture cells. Reagents 1. 1X PBS 2.

By three-color analysis of the transferred cells, the vast majority (90%) were CD8β 1, as was also the case of virtually all BrdU + cells (% of the selected population; Fig. 6 A); % of CD8β 1 cells were CD11c + (in agreement with the percentage observed with CD8α/β 1 CD11c + selected by sorting for repertoire analyses), and all CD11c. Cellular senescence is a state of irreversible growth arrest. In this paper the authors examined whether bleomycin, an agent that causes pulmonary fibrosis, induces the senescence of alveolar epithelial cells. Type II-like alveolar epithelial (A) cells or rat primary type II cells were exposed to bleomycin and then evaluated for markers of cellular senescence. Medical definition of bromodeoxyuridine: a mutagenic analog C9H11O5NBr of thymidine that induces chromosomal breakage especially in heterochromatic regions and has been used to selectively destroy actively dividing cells —abbreviation BUdR. The BrdU antibody can be used for DNA labeling index, evaluation of DNA synthesis and cell proliferation studies. This antibody is biotinylated and thus eliminates the need for a biotinylated secondary antibody. This antibody can be used in all species, including mouse and rat tissues.

studies with bromodeoxyuridine on L-strain mouse cells. by Sop-Chong Kim Download PDF EPUB FB2

BrdU promotes full-chemical induced reprogramming of MEFs. (A) Dose-response effect of BrdU on OKSM-induced reprogramming of MEFs. Starting cell density was 4 MEFs/well (well plate). Summary. The effect of BrdU on oncogenicity and growth characteristics of mouse myeloma cell lines in vitro has been studied.

Oncogenicity was reduced and marked cytotoxicity was observed but globulin synthesis did not appear to be affected by less than lethal by: 1. Alterations in chromosomal morphology and staining affinity were produced in mouse embryo cells after in vivo exposure to mg/kg BrdU on day 10 of gestation.

Treated females were killed either 2, 4, or 6 h after BrdU administration and embryo cells were cultured in a colcemid-containing medium for 2 h.

Mouse embryos exposed to concentrations of 5-bromodeoxyuridine (BUdR) ranging from to 10 /tg/ml in studies with bromodeoxyuridine on L-strain mouse cells.

book for two days from th 8-cele l stag e exhibit a concentration-dependent decrease in the frequency of normal blastocysts and a decrease in average cell number per. Mouse embryos explanted at various stages during neurulation were cultured for 20–28 h in the presence of 25– μg/ml of 5-bromodeoxyuridine (BUdR).

BUdR strongly inhibited closure of the cranial neural tube, which was found to be by: npg r romotes CC eneraton Cell Research ol 25 o 10 Octoer analysis of 4B-CiPSC clone 6 (4B-CiPSC-6) revealed a normal mouse karyotype (40, XY) (Supplementary infor.

Cell cycle duration in equivalent staged neural progenitors from mouse and human was h ± h compared to h ± h, respectively in accordance with cell cycle measurements in other.

5-Bromo-2′-deoxyuridine (BrdU)-retaining cells in the mouse endometrium. (A) At day 7, post-injection, BrdU-retaining cells are present in both epithelial and stromal compartments.(B) In day-old mouse, BrdU was selectively retained (green nuclear staining) in cells at the stromal compartment.(C) In day-old mouse after a chase of 7 weeks and (D) in day-old mouse followed by a chase.

The objectives of the study were to determine whether the 3 different analogues could be used for in vivo studies, and if so, to deter- mine if the CldU incorporation gives the same elevated SCE frequency as is seen in the in vitro studies.

The 3 thymidine analogues, BrdU, IdU, and CldU were obtained from Calbiochem-Behring (LaJolla, CA, U.S.A. Previous studies suggested that BrdU-labeled cells in semi-niferous tubules are preleptotene spermatocytes (Salmand et al. ; Koruji et al. ) and/or A2–A4 spermatogo-nia (Blanco-Rodríguez et al.

; Muñoz-Velasco et al. The combination of the fluorescence techniques—IHC for BrdU, LHC for PNA, and IHC for marker proteins—is.

Bromodeoxyuridine (BrdU) is a halogenated pyrimidine that incorporates into newly synthesized DNA during the S studies with bromodeoxyuridine on L-strain mouse cells. book. BrdU is used ubiquitously in cell birthdating studies and as a means of measuring the proliferative index of various cell populations.

D: pattern of BrdU immunoreactivity in a mouse labelled by BrdU at El6. Nuclei appear immunoreactive in cortical layers II-III. E: photomicrograph of a section similar to that shown in D but counterstained with thionin. istry can be used to map neuronal birthdates, and 1o study neuronal cell kinetics and migration~'~.

In that study, and in a later study (Cameron and McKay, ), it was shown that 50 mg/kg of BrdU does not label all cells in S phase, implying that the optimal dose for BrdU is higher. Another concern of BrdU administration is that cells entering the apoptotic process may take up BrdU, without dividing, before they disappear in apoptosis.

Bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU, BUdR, BrdUrd, broxuridine) is a synthetic nucleoside that is an analog of is commonly used in the detection of proliferating cells in living tissues. 5-Bromodeoxycytidine is deaminated to form BrdU. Methods to analyze cell cycle profiles by flow cytometry using bromodeoxyuridine and propidium iodide staining were also provided, entry of cells in S-phase by [3 H] thymidine incorporation studies, and the analysis of cells in mitosis by staining with antiphospho-H3 antibodies are also provided.

Pancreatic β-cell regeneration, the therapeutic expansion of β-cell number to reverse diabetes, is an important goal.

Replication of differentiated insulin-producing cells is the major source of new β-cells in adult mice and juvenile humans. Nucleoside analogs such as BrdU, which are incorporated into DNA during S-phase, have been widely used to quantify β-cell proliferation.

A method is reported forin vivo bromodeoxyuridine incorporation in mice and its subsequent visualization in kidney by immunohistochemistry. Following formal-sublimate fixation of the kidney, bromodeoxyuridine labelled nuclei were detected in paraffin sections with a monoclonal antibody and visualized by an immunoperoxidase technique.

This rapid and unequivocal method was used to. In contrast to other cells contained within pancreatic preparations, insulin/BrdU copositive cells were fairly rare and comprised only a small portion of total β-cells, even in mice exposed to BrdU for very long periods (Figs. 3 and 5A).

β-Cell proliferation rates averaged ± % per day, far lower than expected given previous data. IIB5 is a Mouse monoclonal IgG1 antibody derived by fusion of SP2/0-Ag14 Mouse myeloma cells with spleen cells from a BALB/c Mouse intraperitoneally immunized with BrdU conjugated to Bovine Serum albumin.

Product Each vial contains ul 1 mg/ml purified monoclonal antibody in PBS containing % sodium azide. Bromodeoxyuridine, variously abbreviated as BrdU, BudR, and BrdUrd, is a halogenated thymidine analog that is permanently integrated into the DNA of dividing cells during DNA synthesis in S phase.

BrdU can be immunocytochemically detected in vitro and in vivo, allowing the identification of cells that were dividing the period of BrdU exposure.

Total counts of BrdU positive cells in the dentate gyrus (i.e., ROI 3) revealed significant differences between ligation injured and sham control brains (Figure 3 and Figure 4).Mean ipsilateral counts in ligation injured brains were ± compared to 32± cells in sham controls and therefore significantly lower (Figure 4A, pBrdU positive cells in non.

DNA replication occurs during S-phase in spermatogonia and preleptotene spermatocytes during spermatogenesis. 5-Bromo-2’-deoxyuridine (BrdU) is incorporated into synthesized DNA and is. In mammalian cells, histones were shown to bind more tightly to 5‐bromouracil‐substituted DNA in vitro than normal DNA containing thymine [[19, 20]].

In this study, we examined whether BrdU induces the expression of genes associated with the destabilization of nucleosome positioning with model plasmids containing the BAR1 gene as a marker. Incubate the tissue in BrdU antisera (, Mouse monoclonal lgG, Dako) overnight at room temperature in a humidified chamber.

Second day Wash the tissue lOX for 10 minutes each with KPBS containing per cent Triton X (Sigma). Expose the tissue sections to horse anti-mouse. limitation in cell killing action, and this theory has been gen erally accepted.

Some investigators (3, 4) have suggested, on the basis of autoradiography with [3H]dThd, that MTX inhibits cell transition at the Gi-S boundary, and recent flow cytometric studies () have also shown Gi-S block on DNA histograms of MTX-treated cells. To study the process by which cells lose BrdU label, we took all cells containing BrdU at day T (all cells in population y 1 and monitored BrdU loss through cell division in the absence of BrdU.

An important goal in diabetes research is to understand the processes that trigger endogenous β-cell proliferation. Hyperglycemia induces β-cell replication, but the mechanism remains debated. A prime candidate is insulin, which acts locally through the insulin receptor. Having previously developed an in vivo mouse hyperglycemia model, we tested whether glucose induces β-cell proliferation.

Wholemount tissue staining was performed to detect BrdU incorporating cells. No cells stained in the control mouse TM without BrdU injection (Fig. 8 A) nor in the BrdU-injected normal mouse TM without cell transplantation (Fig. 8 D), indicating the quiescence of this tissue in adult mouse.

There are also no BrdU-labeled cells in the TM after. IIB5 reacts with BrdU in denatured (single-stranded DNA). The antibody is crossreactive with iododeoxyuridine. It can be used for: 1. Radioimmunocytochemical detection of circulating levels of BrdU 2. Detection of S-phase cells in tissue sections by immunoperoxidase or immunofluorescence method 3.

Detection of S-phase cells in cell suspension 4. Biological Behavior of Mouse CPCs in Vivo. To characterize the pattern of CPC growth and differentiation in the mouse heart, the EGFP lentivirus was injected in proximity of the atrial and apical niches ().The objective was to tag CPCs in situ and identify the clones of differentiated cells derived from the infected CPCs (Fig.

1 A).The recognition of a common integration site in CPCs and. Mouse Anti-Specificity: Anti-BrdU, Clone Bu20a, shows, in common with other monoclonal antibodies of this specificity, weak cross-reactivity with other nucleosides, e.g.

thymidine (3, 4). However, the antibody has been specifically selected with regard to minimal cross-reactivity. See package insert for reference(s).Bromodeoxyuridine (BrdUrd) is used extensively to measure the fraction of proliferating cells in tumors.

Unlike endogenous markers of proliferation such as proliferating cell nuclear antigen (PCNA) and Ki, BrdUrd is exogenously administered and reaches the tumor via vasculature where it must then diffuse throughout the tissue to label S-phase cells. In this study, we examine the dose. Furthermore, BrdU also labels cells that are dying or repairing their DNA.

"BrdU is not a specific marker for new cells," Rakic says. "The caveat is that all dividing cells are labelled, but not.