Cell Cycle Analysis Using In Vivo Staining of DNA-Synthesizing Cells

Páral, Petr; Báječný, Martin; Savvulidi, Filipp; Nečas, E

doi:10.1007/7651_2019_228

Cited by 4 publications

(3 citation statements)

References 10 publications

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“…BrdU is a thymidine analogue, and 7-AAD binds to double stranded DNA by intercalating between base pairs in G-C-rich regions [44]. With this combination, flow cytometric analysis enables the enumeration and characterization of cells that are actively synthesizing DNA (BrdU incorporation) in terms of their cell cycle position (i.e., apoptotic, G0/1, S, or G2/M phase) [45,46].…”

Section: -Bromo-2 -Deoxyuridine (Brdu) Injection and Cell Cycle Analmentioning

confidence: 99%

Comparative Analysis of Age-Related Changes in Lacrimal Glands and Meibomian Glands of a C57BL/6 Male Mouse Model

Yoon

Ryu

Hwang

et al. 2020

IJMS

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It is not known how biological changes in the lacrimal (LGs) and meibomian (MGs) glands contribute to dry eye disease (DED) in a time-dependent manner. In this study, we investigated time-sequenced changes in the inflammation, oxidative stress, and senescence of stem cells in both glands of an aging-related DED mouse model. Eight-week (8W)-, one-year (1Y)-, and two-year (2Y)-old C57BL/6 male mice were used. MG areas of the upper and lower eyelids were analyzed by transillumination meibography imaging. The number of CD45+, 8-OHdG+, Ki-67+, and BrdU+ cells was compared in both glands. Increased corneal staining and decreased tear secretion were observed in aged mice. The MG dropout area increased with aging, and the age-adjusted MG area in lower lids was negatively correlated with the National Eye Institute (NEI) score. Increased CD4+ interferon (IFN)-γ+ cells in LGs were found in both aged mice. An increase in 8-OHdG+ cells in both glands was evident in 2Y-old mice. Reduced Ki-67+ cells, but no change in CD45+ cells, was observed in the MGs of 1Y-old mice. Increased BrdU+ cells were observed in the LGs of aged mice. This suggests that age-dependent DED in C57BL/6 mice is related to inflammation of the LGs, the development of MG atrophy, and oxidative stress in both glands.

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Section: -Bromo-2 -Deoxyuridine (Brdu) Injection and Cell Cycle Analmentioning

confidence: 99%

Comparative Analysis of Age-Related Changes in Lacrimal Glands and Meibomian Glands of a C57BL/6 Male Mouse Model

Yoon

Ryu

Hwang

et al. 2020

IJMS

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show abstract

“…To confirm this finding, flow cytometry was performed. Flow cytometry analysis is a powerful tool utilized for the past decades to investigate cell cycle kinetics and the mechanism of action of bioactive compounds [45][46][47][48][49]. This present study focused on investigating the effect of B. asiatica seed extract on S. cerevisiae cell membrane by staining the cells with calcein AM to distinguish intact cell membrane and propidium iodide identify cells with damaged (raptured) cell membrane.…”

Section: Cell Viability Of Ovarian Carcinoma (A2789) (Live/dead Fluorescence Assay)mentioning

confidence: 99%

Barringtonia asiatica Seed Extract Induces G1 Cell Cycle Arrest in Saccharomyces cerevisiae and Exhibits Cytotoxicity in A2780 Human Ovarian Cancer Cells

Villanueva

Vicera

Leonida

et al. 2021

JPRI

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Barringtonia asiatica, is traditionally used as a medicinal plant in the Philippines; unfortunately, there is limited evidence supporting its anticancer capability. Thus, our study investigated the cytotoxic effect of B. asiatica seed extract against ovarian carcinoma A2780 (ECACC 93112519) and Saccharomyces cerevisiae as a cancer model organism. Seeds of B. asiatica were freeze-dried and extracted with methanol. The seed extract was investigated for its effect on the cell cycle in S. cerevisiae by utilizing yeast budding experiment and imaging flow cytometry. Its cytotoxicity activity was also tested as well as a live/dead assay was conducted against ovarian cancer cells. Findings revealed the anticancer activity of B. asiatica and its capability to induce G1 cell cycle arrest in S. cerevisiae. Thus, B. asiatica seeds may serve as a potential source of natural compounds towards anticancer drug discovery.

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“…2018, Páral et al. 2020). The site of growth inhibition and cell cycle arrest was analyzed using BrdU in MDBK cells (Li 2017).…”

Section: Introductionmentioning

confidence: 98%

Flow cytometry and 5‐ethynyl‐2′‐deoxyuridine (EdU) labeling to detect the cell cycle dynamics of Phaeodactylum tricornutum under light

Zhang

Huang

Zhang

et al. 2022

Journal of Phycology

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Cell cycle studies in plants and algae are highly dependent on reliable methods for detecting cellular DNA replication. With its short growth cycle and ease of genetic transformation, Phaeodactylum tricornutum is an important model organism for the study of pennate diatoms. Here we explored two different methods to detect the cell cycle of P. tricornutum, one using SYBR‐green I to via flow cytometry, and the other using EdU labeling to observe cell cycle changes under fluorescence microscopy. Both EdU labeling fluorescence microscopy and SYBR‐green I staining flow cytometry accurately indicated that the cells of P. tricornutum enter the G2/M phase after 12 h of light exposure. The results indicate that SYBR Green I was an adequate detection method for nuclear DNA quantitation in cells of P. tricornutum using a flow cytometer and without RNase A treatment. In addition, EdU can be applied to P. tricornutum to reliably detect cell proliferation. Besides, Mg‐ProtoIX was able to reverse the cell cycle division inhibition of P. tricornutum and allow the nuclear DNA replication to proceed normally. Taken together, the photoperiodic division time point was clearly identified, which sheds light on the regulation of cell division mechanism in P. tricornutum.

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Cell Cycle Analysis Using In Vivo Staining of DNA-Synthesizing Cells

Cited by 4 publications

References 10 publications

Comparative Analysis of Age-Related Changes in Lacrimal Glands and Meibomian Glands of a C57BL/6 Male Mouse Model

Comparative Analysis of Age-Related Changes in Lacrimal Glands and Meibomian Glands of a C57BL/6 Male Mouse Model

Barringtonia asiatica Seed Extract Induces G1 Cell Cycle Arrest in Saccharomyces cerevisiae and Exhibits Cytotoxicity in A2780 Human Ovarian Cancer Cells

Flow cytometry and 5‐ethynyl‐2′‐deoxyuridine (EdU) labeling to detect the cell cycle dynamics of Phaeodactylum tricornutum under light

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