Stimulating effect of erythropoietin on thymocyte energetics established in vitro with a potential-sensitive fluorescent probe

Morozova, G. I.; Parkhomenko, Tatiana V.; Klitsenko, O. E.; Tomson, V. V.

doi:10.1134/s1990747807040083

Cited by 7 publications

(13 citation statements)

References 4 publications

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“…Hence, EPO is able to exert fast immediate action upon T-lymphoid cells over short incubation terms. Indeed, we have also revealed modulatory eff ect of EPO upon rat thymocytes using a specifi c potential-sensitive chemical probe [9]. EPO was found to exert activating eff ect upon the TLC by changing total transmembrane potential of plasmatic (Δφp) and mitochondrial membranes (Δφm).…”

Section: Resultsmentioning

confidence: 83%

“…We have studied lymphoid cells isolated from thymuses of white Wistar rats (200-300 g), aft er gentle mincing of thymus glands [9]. Th e cells were resuspended in standard Hank's solution at 2 to 3x10 7 cells/mL.…”

Section: Methodsmentioning

confidence: 99%

“…A potential-sensitive fl uorescent probe [4-(p-dimethylaminostyryl)-1-methylpyridinium] (DSM) was used to test the energy potential of cells. DSM was produced and purchased from the Latvian Institute of Organic Synthesis [9]. Th e thymocyte suspensions in Eppendorf-type tubes were pre-incubated with inhibitors for 10…20…40 min at 37°С, then EPO was added at the fi nal concentration of 2U/ml), followed by incubation for 30 min, addition of the DSM probe (1.5 μM), and post-incubated for 20 min.…”

Section: Methodsmentioning

confidence: 99%

“…Fluorescence intensity was manually measured for single cells localized in the vision fi eld, then transformed to a digital signal. Th e numbers of DSM-stained bright mitochondria, looking as intracellular yellow granules, were counted per each single cell (the n m/c values) [9]. Fift y to seventy cells were studied per sample, and the mean fl uorescence intensity was calculated as conventional units (F, arbitrary units).…”

Section: Methodsmentioning

confidence: 99%

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In vitro modifying effect of erythropoietin upon thymic lymphocytes: an inhibitor analysis

Parkhomenko¹,

Tomson²,

Galibin³

2018

CTT

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Section: Resultsmentioning

confidence: 83%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

In vitro modifying effect of erythropoietin upon thymic lymphocytes: an inhibitor analysis

Parkhomenko¹,

Tomson²,

Galibin³

2018

CTT

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“…Transmembrane electric potential is a universal product of the systems of energy coupling. Application of methods using potential sensitive fluorescent probes allows to investigate the response of cells after exposure to various physical and chemical factors [1,4,10,14], and to assess mitochondrial functions and redox potential of hematopoietic cells [8]. Earlier, we have revealed that intensity of fluorescent signal from cationic probe, iodide 2 [p-(dimethylamino)styryl]-1-methylpyridinium (2-Di-1-ASP) is sensitive to changes of energy potential in living cells, including BMC [12].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of energy potential of fresh and stored bone marrow cells using a fluorescent potential-sensitive probe

Parkhomenko¹,

Galibin²,

Verbitskaya³

et al. 2016

CTT

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SummaryBone marrow is a primary source of hematopoietic stem cells in clinical transplantation. Quality of bone marrow grafts is a key factor of their successful in vivo expansion. The aim of our work was to test a semi-quantitative technique for assessment of bone marrow cell viability under strict storage conditions, by means of a fluorescent membrane potential-sensitive 2-Di-1-ASP probe.We have studied 20 samples of normal bone marrow cells (BMC). The cells were placed in a standard storage solution with sodium citrate, citric acid; phosphate salts, dextrose and adenine. Cell counts and viability tests were performed up to 72 hours of incubation. The samples were labeled with 2-Di-1-ASP probe at specified terms. Fluorescence intensity was measured for single nucleated cells, followed by calculating mean fluorescence values and myelokaryocyte numbers. Mitotic indexes were determined both in Giemsa-stained and fluorescent probe-stained cells. Cluster analysis and non-parametric tests were used for statistical evaluation. ResultsInitial cell survival of 80-92% was shown at 3…5 hours of storage, then decreased to 70-75% by the end of incubation. Meanwhile, cell incubation for 3 hours was accompanied by increased fluorescence, in terms of F̃ values, mainly, due to higher proportion of "bright" cell population (>100 arb.units, N F>100 ,%). D (ratio of N F>100 at 3h storage to N F>100 initial) proved to be the most informative parameter, thus enabling us to predict sample-specific differences for the F̃ values at later terms. All BMC samples exhibited increased F, on the account of brighter cell population (N F>100 ), over 3 hours of incubation. This increase correlated with increase in myelokaryocyte counts. An additional cluster analysis allowed us to classify the BMC samples into 3 sub-groups, by their significant inter-group differences for D values and cell number changes. In particular, a number of mitotic cells were detected in BMC populations at 5 to 24 hours of incubation, showing bright stainability with 2-Di-1-ASP probe. We revealed 0.60±0.10% of metaphase cells at initial time point. After 5-h storage, the frequency of mitotic cells increased to 1.4±0.1%; and, after 6-h colchicine treatment, the mitotic index increased to: 1.8±0.1%, thus showing good preservation of dividing cell fraction.In summary, our results have shown sustained, and even increased energy activity using a potential-sensitive probe, and good survival of mitotic cell fraction under strict incubation conditions. Appropriate mechanistic studies of the bone marrow cell preservation and energy balance under the given storage conditions should be performed in future.

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Effect of Glycyrrhizic Acid and Arabinogalactan on the Membrane Potential of Rat Thymocytes Studied by Potential-Sensitive Fluorescent Probe

et al. 2020

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Stimulating effect of erythropoietin on thymocyte energetics established in vitro with a potential-sensitive fluorescent probe

Cited by 7 publications

References 4 publications

In vitro modifying effect of erythropoietin upon thymic lymphocytes: an inhibitor analysis

In vitro modifying effect of erythropoietin upon thymic lymphocytes: an inhibitor analysis

Evaluation of energy potential of fresh and stored bone marrow cells using a fluorescent potential-sensitive probe

Effect of Glycyrrhizic Acid and Arabinogalactan on the Membrane Potential of Rat Thymocytes Studied by Potential-Sensitive Fluorescent Probe

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