Flow Cytometric Analysis of Cell Division by Dilution of CFSE and Related Dyes

2013 ◽  
Vol 64 (1) ◽  
Author(s):  
A. Bruce Lyons ◽  
Stephen J. Blake ◽  
Kathleen V. Doherty
Keyword(s):  
Cell Division ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric

Application of flow cytometric cell cycle analysis to the assessment of condition and growth in larvae of a freshwater teleost Galaxias olidus

2000 ◽  
Vol 57 (4) ◽  
pp. 732-741 ◽  
Author(s):  
Don Bromhead ◽  
John Kalish ◽  
Paul Waring
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Larval Fish ◽  
Fish Larvae ◽  
Flow Cytometric Analysis ◽  
Geographic Location ◽  
Time Of Day ◽  
Flow Cytometric ◽  
Effects Of Temperature ◽  
The Brain

Through its ability to measure cell DNA content, flow cytometric analysis (FCA) is a technique capable of accurately assessing the position of cells in the cell cycle. Using FCA to measure the proportion of dividing and nondividing cells, an index was created that allows the amount of cell division within larval fish tissues to be quantified. To assess the suitability of the cell division index (CDI) as an indicator of growth and condition in fish larvae, analyses were divided into four parts. These examined the effects of temperature, nutrition, time of day, and geographic location on the CDI of brain tissue from Galaxias olidus larvae. The index was sensitive to, firstly, differences in the brain CDI of larvae reared at 12 and 20°C and, secondly, to significant fluctuations in mean brain CDI from larvae sampled over 24 h. FCA also revealed significant differences in the CDI of starving and fed larvae. Overall, this study indicates that FCA may be suitable as an indicator of growth and condition in both laboratory-reared and wild fish larvae.

scholarly journals Actin-based myosin XXI (13) molecular motor is involved in early phase of Leishmania cytokinesis

2021 ◽  
Author(s):  
Rani Bajaj ◽  
Chhitar M. Gupta
Keyword(s):  
Cell Division ◽  
Cell Growth ◽  
Molecular Motor ◽  
Flow Cytometric Analysis ◽  
Microscopic Analysis ◽  
Cell Division Cycle ◽  
Significant Delay ◽  
Initiation Phase ◽  
Flow Cytometric ◽  
Tail Structure

AbstractLeishmania genome encodes for two isoforms of myosin, but only Myosin XXI (Myo21), which is a novel form of myosin in that it contains two ubiquitin associated-like (UBA) domains towards the end of its tail structure, is expressed in both the promastigote and amastigote forms of this protozoan. Earlier studies have shown that in Leishmania promastigotes Myo21 besides localizing throughout the cell body and flagellum, it is prominently localized to the base of the flagellum. It has further been shown that this protein in the promastigotes plays an important role in regulating the cell morphology, motility, flagellum dynamics, growth and intracellular trafficking, As Myo21 depletion has been shown to result in reduced cell growth in culture, we considered it of interest to investigate whether the observed effect of Myo21 on the cell growth is mediated through its possible role in Leishmania cell division cycle. For this, we prepared heterozygous Myo21 mutants of Leishmania promastigotes (Myo21+/−cells) and then analyzed their morphology, growth and cell division cycle, using wild type Leishmania promastigotes (Myo21+/+ cells) as control. The cell division cycle was analyzed by employing flow cytometry and immunofluorescence microscopy. Flow cytometric analysis revealed that the G2/M to G1 phase transition in Myo21+/− cell is significantly delayed, as compared to Myo21+/+ cells. Immunofluorescence confocal microscopic analysis indicated that Myo21+/− cells encountered a significant delay in initiation of cytokinesis, which was mainly due to delay in the flagellar pocket division. Further analysis revealed that actin-based Myo21 motor is essentially required in the initiation phase of Leishmania cytokinesis.

Apoptotic Effects of Melittin on 4T1 Breast Cancer Cell Line is associated with Up Regulation of Mfn1 and Drp1 mRNA Expression

2020 ◽  
Vol 20 (7) ◽  
pp. 790-799 ◽  
Author(s):  
Farnaz D. Moghaddam ◽  
Pejman Mortazavi ◽  
Somayeh Hamedi ◽  
Mohammad Nabiuni ◽  
Nasim H. Roodbari
Keyword(s):  
Breast Cancer ◽  
Mrna Expression ◽  
Hemolytic Activity ◽  
Breast Cancer Cells ◽  
Flow Cytometric Analysis ◽  
Control Group ◽  
Flow Cytometric ◽  
4T1 Cells ◽  
4T1 Breast Cancer ◽  
Apoptotic Effects

Background and Purpose: Melittin, as the main ingredient of honeybee venom, that has shown anticancer properties. The present study aimed at investigating the cytotoxic impacts of melittin on 4T1 breast cancer cells. Methods: Hemolytic activity of different concentrations (0.125, 0.25, 0.5, 1, 2, 4, 8μg/ml) of melittin was assayed and then cytotoxicity of selected concentrations of melittin (2, 4, 8, 16, 32, and 64μg/ml), 2 and 4μg/ml of cisplatin and 0.513, 0.295 and 0.123μg/ml of doxorubicin was evaluated on 4T1 cells using MTT assay. We used Morphological evaluation and flow cytometric analysis was used. Real time PCR was also used to determine mRNA expression of Mfn1 and Drp1 genes. Results: All compounds showed anti-proliferative effects on the tumor cell line with different potencies. Melittin had higher cytotoxicity against 4T1 breast cancer cells (IC50= 32μg/ml-72h) and higher hemolytic activity (HD50= 1μg/ml), as compared to cisplatin and doxorubicin. Mellitin at 16 and 32μg/ml showed apoptotic effects on 4T1 cells according to the flow cytometric analysis. The Real time PCR analysis of Drp1 and Mfn1 expression in cells treated with 16μg/ml of melittin revealed an up-regulation in Drp1 and Mfn1 genes mRNA expression in comparison with control group. Treatment with 32μg/ml of melittin was also associated with a rise in mRNA expression of Drp1 and Mfn1 as compared to the control group. Conclusion: The results of this study showed that melittin has anticancer effects on 4T1 cell lines in a dose and time dependent manner and can be a good candidate for further research on breast cancer treatment.

scholarly journals Flow cytometric analysis of T lymphocytes and cytokines in aqueous humor of patients with varicella zoster virus-mediated acute retinal necrosis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Hao Kang ◽  
Yunbo Wei ◽  
Ming Liu ◽  
Di Yu ◽  
Yong Tao
Keyword(s):  
T Lymphocytes ◽  
Varicella Zoster Virus ◽  
Aqueous Humor ◽  
T Lymphocyte ◽  
Lymphocyte Subsets ◽  
Flow Cytometric Analysis ◽  
T Lymphocyte Subsets ◽  
Varicella Zoster ◽  
Flow Cytometric ◽  
Cd8 T Lymphocytes

Abstract Background The purpose of this study is to investigate the aqueous humor (AH) T lymphocyte subsets and cytokines of acute retinal necrosis (ARN) to elucidate the immunologic inflammatory features of this disorder. Methods Three patients with ARN infected with varicella zoster virus (VZV) who underwent multiple intravitreal injections of ganciclovir were enrolled in this study. The control group consisted of four non-infectious patients with acute anterior uveitis (AAU). Flow cytometric analysis was performed on the lymphocyte subsets from the AH and peripheral blood (PB) samples during the active phase of intraocular inflammation. Five inflammatory cytokines were measured in each AH sample and various clinical characteristics were also assessed. Results VZV deoxyribonucleic acid (DNA) was detected by real-time polymerase chain reaction (PCR) in AH from all the ARN patients, who showed higher CD8+ T lymphocytes population in AH than the AAU patients (p = 0.006). CD4/CD8 ratios of T lymphocytes and the percentage of CD8 + CD25+ T lymphocytes in AH were significantly lower in ARN than in AAU (p = 0.006; p = 0.012). In the ARN patients, the percentages of CD4+ and CD8+ T lymphocytes in AH were higher than those found in PB. The percentage of CD4 + CD25+ T lymphocytes in AH was significantly higher than the proportion in PB in the AAU patients (p = 0.001). Immunoregulatory cytokine Interleukin-10 in AH was significantly elevated in the ARN patients in comparison with the case of the AAU patients (p = 0.036). In ARN, the copy number of VZV DNA in AH positively correlated with the percentage of CD8+ T lymphocytes in AH and negatively correlated with the CD4/CD8 ratio in AH during the course of disease treatment (p = 0.009, r = 0.92; p = 0.039, r = − 0.834). Conclusion The ARN patients caused by VZV had different intraocular T lymphocyte subsets and cytokines profile than those of the non-infectious patients. High percentages of CD8+ T lymphocytes and low CD4/CD8 T cell ratios may be a potential biomarker for diagnosis of viral-infectious uveitis. T lymphocytes examination at the inflammatory sites has the potential to become a useful research tool for differentiating viral and non-viral uveitis.

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