Quantitative multiwell myeloid differentiation assay using dichlorodihydrofluorescein diacetate (H2DCF-DA) or dihydrorhodamine 123 (H2R123)

Trayner, Ian D.; Rayner, Anne; Freeman, Gwenyth; Farzaneh, Farzin

doi:10.1016/0022-1759(95)00152-z

Cited by 54 publications

(26 citation statements)

References 24 publications

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“…Cells were stimulated with PMA to activate NADPH oxidase and incubated in the presence of CM-H 2 DCFDA. CM-H 2 DCFDA passively diffuses into cells and, following cleavage of acetate groups by intracellular esterases, oxidation by superoxide anion/hydrogen peroxide yields a fluorescent adduct that is trapped inside the cell, facilitating its detection by flow cytometry (33,35). Cells treated for 5 days with 1 Amol/L RA or 1 Amol/L D 3 , which cause differentiation of HL-60 cells to granulocytes and monocytes, respectively, were used as positive controls.…”

Section: Resultsmentioning

confidence: 99%

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Cholesterol Starvation Induces Differentiation of Human Leukemia HL-60 Cells

et al. 2007

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Cholesterol metabolism is particularly active in malignant, proliferative cells, whereas cholesterol starvation has been shown to inhibit cell proliferation. Inhibition of enzymes involved in cholesterol biosynthesis at steps before the formation of 7-dehydrocholesterol has been shown to selectively affect cell cycle progression from G 2 phase in human promyelocytic HL-60 cells. In the present work, we explored whether cholesterol starvation by culture in cholesterol-free medium and treatment with different distal cholesterol biosynthesis inhibitors induces differentiation of HL-60 cells. Treatment with SKF 104976, an inhibitor of lanosterol 14-A demethylase, or with zaragozic acid, which inhibits squalene synthase, caused morphologic changes alongside respiratory burst activity and expression of cluster of differentiation antigen 11c (CD11c) but not cluster of differentiation antigen 14. These effects were comparable to those produced by all-trans retinoic acid, which induces HL-60 cells to differentiate following a granulocyte lineage. In contrast, they differed from those produced by vitamin D 3 , which promotes monocyte differentiation. The specificity of the response was confirmed by addition of cholesterol to the culture medium. Treatment with PD 98059, an inhibitor of extracellular signal-regulated kinase, abolished both the activation of NADPH oxidase and the expression of the CD11c marker. In sharp contrast, BM 15766, which inhibits sterol # 7 -reductase, failed to induce differentiation or arrest cell proliferation. These results show that changes in the sterol composition may trigger a differentiation response and highlight the potential of cholesterol pathway inhibition as a possible tool for use in cancer therapy. [Cancer Res 2007;67(7):3379-86]

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Section: Resultsmentioning

confidence: 99%

“…The respiratory burst was detected by oxidation of the fluorescent probe CM-H 2 DCFDA, as described elsewhere (33). Cells (5 Â 10 CD marker expression.…”

Section: Methodsmentioning

confidence: 99%

Cholesterol Starvation Induces Differentiation of Human Leukemia HL-60 Cells

et al. 2007

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“…The ROS assay was modified from previously described techniques (Rosenkranz et al, 1992;Trayner et al, 1995;Miller et al, 1996;Swerdlow et al, 1996). Cells were plated at a density of 100,000 cells/well in 96-well tissue culture plates and allowed to grow for 24 hr in DMEM supplemented with 10% fetal bovine serum.…”

Section: Methodsmentioning

confidence: 99%

Calcium Homeostasis and Reactive Oxygen Species Production in Cells Transformed by Mitochondria from Individuals with Sporadic Alzheimer’s Disease

Sheehan

Swerdlow

Miller³

et al. 1997

J. Neurosci.

241

140

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Alzheimer's disease (AD) is associated with defects in mitochondrial function. Mitochondrial-based disturbances in calcium homeostasis, reactive oxygen species (ROS) generation, and amyloid metabolism have been implicated in the pathophysiology of sporadic AD. The cellular consequences of mitochondrial dysfunction, however, are not known. To examine these consequences, mitochondrially transformed cells (cybrids) were created from AD patients or disease-free controls. Mitochondria from platelets were fused to 0 cells created by depleting the human neuroblastoma line SH-SY5Y of its mitochondrial DNA (mtDNA). AD cybrids demonstrated a 52% decrease in electron transport chain (ETC) complex IV activity but no difference in complex I activity compared with control cybrids or SH-SY5Y cells. This mitochondrial dysfunction suggests a transferable mtDNA defect associated with AD. ROS generation was elevated in the AD cybrids. AD cybrids also displayed an increased basal cytosolic calcium concentration and enhanced sensitivity to inositol-1,4,5-triphosphate (InsP 3 )-mediated release. Furthermore, they recovered more slowly from an elevation in cytosolic calcium induced by the InsP 3 agonist carbachol. Mitochondrial calcium buffering plays a major role after this type of perturbation. ␤-amyloid (25-35) peptide delayed the initiation of calcium recovery to a carbachol challenge and slowed the recovery rate. Nerve growth factor reduced the carbachol-induced maximum and moderated the recovery kinetics. Succinate increased ETC activity and partially restored the AD cybrid recovery rate. These subtle alterations in calcium homeostasis and ROS generation might lead to increased susceptibility to cell death under circumstances not ordinarily toxic.

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“…Fig. 1B; Ubezio and Civoli, 1994;Trayner et al, 1995;Allan and Fluhr, 1997). Oxidation of this dye by O 2 Ϫ or by H 2 O 2 catalyzed by peroxidase produces a fluorescent and cell-permeable product, 2Ј,7Ј-dichlorofluorescein, that can be detected in the medium (Ubezio and Civoli, 1994).…”

Section: Generation Of Ros In Differentiating Cotton Fibersmentioning

confidence: 99%

The Involvement of Hydrogen Peroxide in the Differentiation of Secondary Walls in Cotton Fibers1

et al. 1999

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The signals that control the developmental switch from primary to secondary wall synthesis in higher plants are not known. This transition is characterized by a cessation of synthesis of polymers unique to the primary cell wall accompanied by enhanced rates of cellulose deposition and induction of synthesis of specific secondary wall matrix polysaccharides and lignin. The developing fibers of cotton (Gossypium hirsutum) and tracheary elements differentiated from isolated mesophyll cells of zinnia have emerged as two good model systems for studying this transition. In the zinnia system the mesophyll cells are cultured in noninductive medium and differentiation is induced by changes in hormonal balance. Specialized, localized regions of secondary wall formation unique to these tracheary elements have been shown to contain large amounts of cellulose, xylan, and lignin (for reviews, see Fukuda, 1991Fukuda, , 1996. The developing cotton fiber is unique in that the secondary wall consists of nearly pure cellulose and is devoid of hemicellulose and lignin (for reviews of cotton fiber development, see Basra and Malik, 1984;Ryser, 1985). Furthermore, development occurs synchronously for nearly all fibers within a boll, with the transition to secondary wall formation beginning abruptly in varieties of cotton at about 14 to 16 DPA, which is a few days prior to the cessation of fiber elongation (Meinert and Delmer, 1977).Rates of secondary wall cellulose synthesis peak at about 24 DPA; the fibers mature and die sometime after 40 DPA, presumably by a process of programmed cell death. During the transition the rate of cellulose synthesis increases abruptly to about 100-fold (Meinert and Delmer, 1977), and recent evidence indicates that genes that most likely encode the catalytic subunit of cellulose synthase are also strongly induced at this time (Pear et al., 1996). During xylogenesis there is also evidence that expression of genes involved in synthesis of hemicellulose (Bolwell and Northcote, 1981) and lignin (Fukuda, 1991) also undergo induction. The transition in both cotton fibers and tracheary elements is also characterized by a reorganization of the cytoskeleton that directs the specific patterns of cellulose deposition (Seagull, 1990; Fukuda, 1991 Fukuda, , 1996.Our interest in the events regulating this transition was stimulated by our recent characterization of genes that encode two small GTPases of the Rho subfamily in cotton, named Rac13 and Rac9. Rac13 in particular shows highly induced expression at the transition from primary to secondary wall synthesis (Delmer et al., 1995). In animals Rac proteins function in several possibly related signal transduction pathways. Rac has been shown to be involved in regulation of reorganization of the actin cytoskeleton (Symons, 1996) and it plays another role in leukocytes as a specific activator of the plasma membrane NADPH oxidase (Freeman et al., 1996). Activation of the NADPH oxidase leads to generation of the oxidative burst, which serves as a defense against pathoge...

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Quantitative multiwell myeloid differentiation assay using dichlorodihydrofluorescein diacetate (H2DCF-DA) or dihydrorhodamine 123 (H2R123)

Cited by 54 publications

References 24 publications

Cholesterol Starvation Induces Differentiation of Human Leukemia HL-60 Cells

Cholesterol Starvation Induces Differentiation of Human Leukemia HL-60 Cells

Calcium Homeostasis and Reactive Oxygen Species Production in Cells Transformed by Mitochondria from Individuals with Sporadic Alzheimer’s Disease

The Involvement of Hydrogen Peroxide in the Differentiation of Secondary Walls in Cotton Fibers1

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