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1.
Arch Virol ; 165(1): 207-214, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31776677

RESUMO

Bovine leukemia virus (BLV) infects cattle worldwide and causes B-cell lymphoma in cattle. BLV has been identified in human breast and lung cancer and in blood, but the association of BLV and human cancer is controversial. In this study, we investigated the existence of BLV in 145 Japanese human blood cell lines and 54 human cancer cell lines, using a new highly sensitive PCR assay that can amplify even one copy of BLV using LTR primers different from those in previous studies on BLV provirus in breast cancer. All samples were found negative for BLV provirus, suggesting that BLV is unlikely to infect humans.


Assuntos
Células Sanguíneas/virologia , Linhagem Celular Tumoral/virologia , Vírus da Leucemia Bovina/isolamento & purificação , Zoonoses/diagnóstico , Adulto , Idoso , Animais , Células Sanguíneas/citologia , Linhagem Celular , Linhagem Celular Tumoral/citologia , Feminino , Humanos , Japão , Vírus da Leucemia Bovina/genética , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade , Sequências Repetidas Terminais , Adulto Jovem , Zoonoses/virologia
2.
Tissue Cell ; 59: 10-17, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31383284

RESUMO

Domestic Pigeon (Columba livia) is one of few domesticated birds with an important economic value. In this study, a comprehensive investigation on the morphology and cytochemical patterns of peripheral blood cells in domestic pigeons were conducted by using wright's and various cytochemical staining techniques including periodic acid-Schiff(PAS), sudan black B(SBB), peroxidase(POX), alkaline phosphatase(ALP), acid phosphatase(ACP), chloroacetic acid AS-D naphthol esterase(AS-D) and α-naphthol acetate esterase(α-NAE) staining. Besides erythrocytes and thrombocytes, five types of leukocytes were identified: heterophils, eosinophils, basophils, lymphocytes and monocytes. Lymphocytes were the most abundant leukocytes, followed by heterophils, eosinophils, monocytes; basophils were the fewest. Erythrocytes and thrombocytes were positive for PAS, and negative for all the other cytochemical staining. Heterophils and eosinophils exhibited positive to all cytochemical staining except for α-NAE. Basophils exhibited strongly positive for POX and AS-D, positive for PAS and ALP, while negative for SBB, ACP and α-NAE staining. Monocytes exhibited positive for PAS and α-NAE, and weakly positive for ACP, while negative for SBB, POX, ALP and AS-D staining. Lymphocytes showed positive for PAS and ACP, weakly positive for AS-D, while negative for SBB, POX, ALP and α-NAE staining. Our results add up knowledge about the domestic pigeon blood cells.


Assuntos
Células Sanguíneas/citologia , Células Sanguíneas/metabolismo , Columbidae/metabolismo , Animais , Coloração e Rotulagem
4.
J Environ Sci Health B ; 54(10): 866-874, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31258003

RESUMO

Mikania glomerata Sprengel, popularly known as "guaco," is used in Brazilian folk medicine for several inflammatory and allergic conditions. Besides, the popular use "guaco" is indicated by the Brazilian Ministry of Health as a safe and effective herbal medicine. The biological activity of M. glomerata extracts is due to the presence of the coumarins, a large family of phenolic substances found in plants and is made of fused benzene and α-pyrone rings. Considering that there are few data on the biological effects of the extracts of M. glomerata, mainly in genetic level, this work aims to evaluate, in vitro, the genotoxicity and coumarin production in M. glomerata in conventional and organic growing. The data showed that the organic culture system showed double the concentration of coumarin being significantly more productive than the conventional system. Besides, the results of comet assay suggest that extracts of M. glomerata cultivated in a conventional system was genotoxic, increased DNA damage levels while the organic extracts seem to have antigenotoxic effect possibly due to the concentration of coumarins. Additional biochemical investigations are necessary to elucidate the mechanisms of action of M. glomerata extracts, which were found to have a role in protection against DNA damage.


Assuntos
Agricultura/métodos , Cumarínicos/metabolismo , Mikania/metabolismo , Extratos Vegetais/toxicidade , Plantas Medicinais/metabolismo , Células Sanguíneas/citologia , Células Sanguíneas/efeitos dos fármacos , Brasil , Sobrevivência Celular/efeitos dos fármacos , Cumarínicos/toxicidade , Dano ao DNA/efeitos dos fármacos , Humanos , Mikania/química , Testes de Mutagenicidade , Agricultura Orgânica/métodos , Extratos Vegetais/análise , Extratos Vegetais/química
5.
Cells ; 8(7)2019 07 13.
Artigo em Inglês | MEDLINE | ID: mdl-31337010

RESUMO

Commonly, circulating tumor cells (CTCs) are described as source of metastasis in cancer patients. However, in this process cancer cells of the primary tumor site need to survive the physical and biological challenges in the blood stream before leaving the circulation to become the seed of a new metastatic site in distant parenchyma. Most of the CTCs released in the blood stream will not resist those challenges and will consequently fail to induce metastasis. A few of them, however, interact closely with other blood cells, such as neutrophils, platelets, and/or macrophages to survive in the blood stream. Recent studies demonstrated that the interaction and modulation of the blood microenvironment by CTCs is pivotal for the development of new metastasis, making it an interesting target for potential novel treatment strategies. This review will discuss the recent research on the processes in the blood microenvironment with CTCs and will outline currently investigated treatment strategies.


Assuntos
Células Sanguíneas/metabolismo , Células Neoplásicas Circulantes/metabolismo , Microambiente Tumoral , Células Sanguíneas/citologia , Comunicação Celular , Humanos
6.
Elife ; 82019 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-31237234

RESUMO

Our results highlight for the first time that a significant proportion of cell doublets in flow cytometry, previously believed to be the result of technical artifacts and thus ignored in data acquisition and analysis, are the result of biological interaction between immune cells. In particular, we show that cell:cell doublets pairing a T cell and a monocyte can be directly isolated from human blood, and high resolution microscopy shows polarized distribution of LFA1/ICAM1 in many doublets, suggesting in vivo formation. Intriguingly, T cell-monocyte complex frequency and phenotype fluctuate with the onset of immune perturbations such as infection or immunization, reflecting expected polarization of immune responses. Overall these data suggest that cell doublets reflecting T cell-monocyte in vivo immune interactions can be detected in human blood and that the common approach in flow cytometry to avoid studying cell:cell complexes should be re-visited.


Assuntos
Células Sanguíneas/citologia , Adesão Celular , Monócitos/citologia , Monócitos/imunologia , Linfócitos T/citologia , Linfócitos T/imunologia , Citometria de Fluxo , Humanos , Microscopia
7.
PLoS One ; 14(5): e0216602, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31075112

RESUMO

Endothelial dysfunction (ED) is part of the first steps in the development of cardiovascular diseases (CVD). Growth Differentiation Factor 15 (GDF15) is a cytokine belonging to the Transforming Growth Factor ß superfamily and its expression is increased both during ED and in CVD. Because high blood levels of GDF15 have been reported during ED, we hypothesized that GDF15 could be produced by endothelial cells in response to a vascular stress, possibly to attenuate endothelial function loss. Since senescence is mainly involved in both vascular stress and endothelial function loss, we used Endothelial Colony Forming Cells generated from adult blood (AB-ECFCs) as a model of endothelial cells to investigate GDF15 expression during cellular senescence. Then, we analyzed the potential role of GDF15 in AB-ECFC functions and senescence. When AB-ECFCs become senescent, they secrete increased levels of GDF15. We investigated GDF15 paracrine effects on non-senescent AB-ECFCs and showed that GDF15 enhanced proliferation, migration, NO production and activated several signaling pathways including AKT, ERK1/2 and SMAD2 without triggering any oxidative stress. Taken together, our results suggest that GDF15 production by senescent AB-ECFCs could act in a paracrine manner on non-senescent AB-ECFCs, and that this interaction could be beneficial to its model cells. Therefore, GDF15 could play a beneficial role in a dysfunctional vascular system as previously reported in patients with CVD, by limiting ED related to vascular stress occurring in these diseases.


Assuntos
Células Sanguíneas/citologia , Células Endoteliais/citologia , Fator 15 de Diferenciação de Crescimento/genética , Fator 15 de Diferenciação de Crescimento/metabolismo , Adulto , Idoso , Células Sanguíneas/metabolismo , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Senescência Celular , Células Endoteliais/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Óxido Nítrico/metabolismo , Estresse Oxidativo , Transdução de Sinais , Regulação para Cima , Adulto Jovem
8.
Nucleic Acids Res ; 47(11): e65, 2019 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-30941409

RESUMO

The chromosome conformation capture (3C) technique and its variants have been employed to reveal the existence of a hierarchy of structures in three-dimensional (3D) chromosomal architecture, including compartments, topologically associating domains (TADs), sub-TADs and chromatin loops. However, existing methods for domain detection were only designed based on symmetric Hi-C maps, ignoring long-range interaction structures between domains. To this end, we proposed a generic and efficient method to identify multi-scale topological domains (MSTD), including cis- and trans-interacting regions, from a variety of 3D genomic datasets. We first applied MSTD to detect promoter-anchored interaction domains (PADs) from promoter capture Hi-C datasets across 17 primary blood cell types. The boundaries of PADs are significantly enriched with one or the combination of multiple epigenetic factors. Moreover, PADs between functionally similar cell types are significantly conserved in terms of domain regions and expression states. Cell type-specific PADs involve in distinct cell type-specific activities and regulatory events by dynamic interactions within them. We also employed MSTD to define multi-scale domains from typical symmetric Hi-C datasets and illustrated its distinct superiority to the-state-of-art methods in terms of accuracy, flexibility and efficiency.


Assuntos
Algoritmos , Mapeamento Cromossômico/métodos , Biologia Computacional/métodos , Genômica/métodos , Regiões Promotoras Genéticas , Animais , Sítios de Ligação , Células Sanguíneas/citologia , Células Cultivadas , Cromatina , Elementos Facilitadores Genéticos , Epigênese Genética , Humanos , Camundongos , Polimorfismo de Nucleotídeo Único
9.
ACS Appl Mater Interfaces ; 11(12): 11157-11166, 2019 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-30869853

RESUMO

It is well known that tumors have an acidic pH microenvironment and contain a high content of hydrogen peroxide (H2O2). These features of the tumor microenvironment may provide physiochemical conditions that are suitable for selective tumor therapy and recognition. Here, for the first time, we demonstrate that a type of graphene oxide nanoparticle (N-GO) can exhibit peroxidase-like activities (i.e., can increase the levels of reactive oxygen species (ROS)) under acidic conditions and catalyze the conversion of H2O2 to ROS-hydroxyl radicals (HO·) in the acidic microenvironment in Hela tumors. The concentrated and highly toxic HO· can then trigger necrosis of tumor cells. In the microenvironment of normal tissues, which has a neutral pH and low levels of H2O2, N-GOs exhibit catalase-like activity (scavenge ROS) that splits H2O2 into O2 and water (H2O), leaving normal cells unharmed. In the recognition of tumors, an inherent redox characteristic of dopamine is that it oxidizes to form dopamine-quinine under neutral (pH 7.4) conditions, quenching the fluorescence of N-GOs; however, this characteristic has no effect on the fluorescence of N-GOs in an acidic (pH 6.0) medium. This pH-controlled response provides an active targeting strategy for the diagnostic recognition of tumor cells. Our current work demonstrates that nanocatalytic N-GOs in an acidic and high-H2O2 tumor microenvironment can provide novel benefits that can reduce drug resistance, minimize side effects on normal tissues, improve antitumor efficacy, and offer good biocompatibility for tumor selective therapeutics and specific recognition.


Assuntos
Grafite/química , Peróxido de Hidrogênio/química , Nanopartículas/química , Animais , Células Sanguíneas/citologia , Células Sanguíneas/metabolismo , Catalase/química , Catalase/metabolismo , Catálise , Sobrevivência Celular/efeitos dos fármacos , Dopamina/química , Feminino , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Camundongos , Camundongos Endogâmicos BALB C , Nanopartículas/metabolismo , Nanopartículas/uso terapêutico , Nanopartículas/toxicidade , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Espécies Reativas de Oxigênio/metabolismo , Transplante Heterólogo , Microambiente Tumoral
10.
Nutrients ; 11(3)2019 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-30866565

RESUMO

There is limited evidence from epidemiological studies for the inflammatory or anti-inflammatory properties of fatty acids in blood cell membranes. Therefore, this study examined associations between baseline (n = 282) and 1-year (n = 143) changes in the levels of fatty acids in blood cell membranes with circulating inflammatory markers in older adults at high cardiovascular risk. The data for this cross-sectional analysis was obtained from a case-control study within the PREDIMED study. Linear regression with elastic net penalty was applied to test associations between measured fatty acids and inflammatory markers. Several fatty acids were associated with interferon-γ (IFNγ) and interleukins (ILs) IL-6, IL-8, and IL-10 at baseline and additionally also with IL-1b at 1 year. Omega-6 fatty acids were consistently positively associated with pro-inflammatory IL-6 and IL-8 at baseline. Omega-3 fatty acids including C20:5n3 and C18:3n3 were negatively associated with IFN-γ at 1 year. It is interesting to note that the cis and trans forms of C16:1n7 at 1 year were oppositely associated with the inflammatory markers. C16:1n7trans was negatively associated with IFN-γ, IL-6, IL-8, IL-10, and IL-1b, whereas C16:1n7cis was positively associated with IL-1b. This study adds to the growing body of evidence suggesting potential differences in inflammatory or anti-inflammatory properties of fatty acids in blood cell membranes.


Assuntos
Células Sanguíneas/citologia , Membrana Celular/química , Ácidos Graxos/análise , Inflamação/metabolismo , Idoso , Idoso de 80 Anos ou mais , Células Sanguíneas/química , Estudos de Casos e Controles , Estudos Transversais , Citocinas/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
11.
Mater Sci Eng C Mater Biol Appl ; 99: 491-504, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30889724

RESUMO

Polyurethane (PU) with three different functional groups: carboxyl, hydroxyl and sulphonyl group on its molecular structure were synthesised in this work. The synthesised material suppresses blood clotting and exhibits anticoagulant characteristics due to the presence of the important anionic groups. The synthesised PU was blended with polyethersulphone (PES) and fabricated into flat-sheet membrane to study the physico-chemical and biocompatibility properties of the PES membrane for blood purification application. PES-PU flat-sheet membranes were fabricated via the dry-wet phase separation technique. Different loading of PU (0, 1, 2, 3, 4, and 5%) blended with PES was studied and compared. Based on the in-vitro biocompatibility analysis of the membrane, it can be suggested that the membrane incorporated with PU has better anticoagulant properties compared to the pristine PES membrane. PU incorporation prolonged the clotting time, decreased the formation of thrombin, decreased soluble complement component 3a (C3a) generation and suppressed platelet adhesion and aggregation. The anionic groups on the membrane surface might bind to coagulation factors (antithrombin) and the calcium ions, Ca2+ and thus improve anticoagulant ability. Based on both physico-chemical and in-vitro studied, 4% loading of PU is the optimum loading for incorporation with PES membrane. These results suggested that the blended PES-PU membranes with good haemocompatibility allowed practical application in the field of blood purification.


Assuntos
Células Sanguíneas/citologia , Separação Celular/métodos , Membranas Artificiais , Polímeros/síntese química , Poliuretanos/síntese química , Sulfonas/síntese química , Coagulação Sanguínea , Ativação do Complemento , Complemento C3a/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Microscopia de Força Atômica , Adesividade Plaquetária , Polímeros/química , Poliuretanos/química , Espectroscopia de Prótons por Ressonância Magnética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectroscopia de Infravermelho com Transformada de Fourier , Sulfonas/química , Propriedades de Superfície , Temperatura Ambiente , Trombose/patologia
12.
Dev Cell ; 49(1): 118-129.e7, 2019 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-30827895

RESUMO

The nature of cell-state transitions during the transit-amplifying phases of many developmental processes-hematopoiesis in particular-is unclear. Here, we use single-cell RNA sequencing to demonstrate a continuum of transcriptomic states in committed transit-amplifying erythropoietic progenitors, which correlates with a continuum of proliferative potentials in these cells. We show that glucocorticoids enhance erythrocyte production by slowing the rate of progression through this developmental continuum of transit-amplifying progenitors, permitting more cell divisions prior to terminal erythroid differentiation. Mechanistically, glucocorticoids prolong expression of genes that antagonize and slow induction of genes that drive terminal erythroid differentiation. Erythroid progenitor daughter cell pairs have similar transcriptomes with or without glucocorticoid stimulation, indicating largely symmetric cell division. Thus, the rate of progression along a developmental continuum dictates the absolute number of erythroid cells generated from each transit-amplifying progenitor, suggesting a paradigm for regulating the total output of differentiated cells in numerous other developmental processes.


Assuntos
Células Sanguíneas/metabolismo , Proliferação de Células/genética , Células Precursoras Eritroides/metabolismo , Hematopoese/genética , Animais , Células Sanguíneas/citologia , Diferenciação Celular/genética , Divisão Celular/genética , Células Cultivadas , Eritrócitos/citologia , Eritrócitos/metabolismo , Células Eritroides/citologia , Células Eritroides/metabolismo , Células Precursoras Eritroides/citologia , Eritropoese/genética , Glucocorticoides/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Camundongos , Análise de Célula Única/métodos , Transcriptoma/genética
13.
Methods Mol Biol ; 1953: 253-268, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30912027

RESUMO

Flow cytometry enables the measurement of single cells in a flowing system. Heterogeneous mixtures of cells or particles can be analyzed with respect to their morphology, surface and intracellular protein expression, DNA content, and cellular physiology at high speed and purity. A series of key technical developments and improvements in flow cytometry hardware, software, and dye chemistry made it possible to measure more than 20 parameters simultaneously. Here, we provide a stepwise protocol for the preparation of single cell suspension samples from different murine lymphoid or tumor tissues and a detailed description of a 17-color polychromatic flow cytometry analysis of tumor-infiltrating leukocytes.


Assuntos
Citometria de Fluxo/métodos , Imunofenotipagem/métodos , Animais , Células Sanguíneas/citologia , Células Sanguíneas/imunologia , Células da Medula Óssea/citologia , Células da Medula Óssea/imunologia , Separação Celular/métodos , Linfonodos/citologia , Linfonodos/imunologia , Linfócitos do Interstício Tumoral/citologia , Linfócitos do Interstício Tumoral/imunologia , Camundongos , Neoplasias/imunologia , Análise de Célula Única/métodos , Baço/citologia , Baço/imunologia
14.
Electrophoresis ; 40(10): 1486-1493, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30740752

RESUMO

Detection and analysis of circulating tumor cells (CTCs) have emerged as a promising way to diagnose cancer, study its cellular mechanism, and test or develop potential treatments. However, the rarity of CTCs among peripheral blood cells is a big challenge toward CTC detection. In addition, in cases where there is similar size range between certain types of CTCs (e.g. breast cancer cells) and white blood cells (WBCs), high-resolution techniques are needed. In the present work, we propose a deterministic dielectrophoresis (DEP) method that combines the concept of deterministic lateral displacement (DLD) and insulator-based dielectrophoresis (iDEP) techniques that rely on physical markers such as size and dielectric properties to differentiate different type of cells. The proposed deterministic DEP technology takes advantage of frequency-controlled AC electric field for continuous separation of CTCs from peripheral blood cells. Utilizing numerical modeling, different aspects of coupled DLD-DEP design such as the required applied voltages, velocities, and geometrical parameters of DLD arrays of microposts are investigated. Regarding the inevitable difference and uncertainty ranges for the reported crossover frequencies of cells, a comprehensive analysis is conducted on applied electric field frequency as design's determinant factor. Deterministic DEP design provides continuous sorting of CTCs from WBCs even with similar size and has the future potential for high throughput and efficiency.


Assuntos
Células Sanguíneas/patologia , Separação Celular/métodos , Eletroforese/instrumentação , Eletroforese/métodos , Células Neoplásicas Circulantes/patologia , Células Sanguíneas/química , Células Sanguíneas/citologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Separação Celular/instrumentação , Desenho de Equipamento , Feminino , Humanos , Leucócitos/química , Leucócitos/citologia , Leucócitos/patologia , Células Neoplásicas Circulantes/química
15.
Angiology ; 70(8): 711-718, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30773906

RESUMO

Major advances in coronary interventional techniques and pharmacotherapy as well as the use of drug-eluting stents (DESs) have considerably reduced the risk of in-stent restenosis (ISR). However, ISR remains a major clinical challenge. Inflammation and platelet activation are important processes that underlie the pathophysiology of ISR. Parameters related to blood cells, entailing both cell count and morphology, are useful markers of the inflammatory response and platelet activation in clinical practice. Recent studies have highlighted several new combined or derived parameters related to blood cells that independently predict ISR after DES implantation. The neutrophil/lymphocyte ratio, an inflammatory marker, is regarded as a predictor of the risk of ISR and the stability of atherosclerotic plaques. The mean platelet volume, a widely used platelet activation parameter, has been shown to be a predictor of the risk of ISR and the efficacy of antiplatelet therapy. Other markers considered include the platelet/lymphocyte ratio, red blood cell distribution width, and platelet distribution width. This review provides an overview of these parameters that may help stratify the risk of coronary angiographic and clinical outcomes related to ISR.


Assuntos
Biomarcadores/sangue , Células Sanguíneas/citologia , Reestenose Coronária/terapia , Intervenção Coronária Percutânea , Stents Farmacológicos , Humanos , Intervenção Coronária Percutânea/métodos , Fatores de Risco
16.
Nat Commun ; 10(1): 415, 2019 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-30679420

RESUMO

In life sciences, the material properties of suspended cells have attained significance close to that of fluorescent markers but with the advantage of label-free and unbiased sample characterization. Until recently, cell rheological measurements were either limited by acquisition throughput, excessive post processing, or low-throughput real-time analysis. Real-time deformability cytometry expanded the application of mechanical cell assays to fast on-the-fly phenotyping of large sample sizes, but has been restricted to single material parameters as the Young's modulus. Here, we introduce dynamic real-time deformability cytometry for comprehensive cell rheological measurements at up to 100 cells per second. Utilizing Fourier decomposition, our microfluidic method is able to disentangle cell response to complex hydrodynamic stress distributions and to determine viscoelastic parameters independent of cell shape. We demonstrate the application of our technology for peripheral blood cells in whole blood samples including the discrimination of B- and CD4+ T-lymphocytes by cell rheological properties.


Assuntos
Forma Celular , Citometria de Fluxo/métodos , Microfluídica/métodos , Reologia/métodos , Análise de Célula Única/métodos , Células Sanguíneas/citologia , Forma Celular/efeitos dos fármacos , Citocalasina D/farmacologia , Elasticidade , Células HL-60/efeitos dos fármacos , Humanos , Hidrodinâmica , Modelos Biológicos , Fenótipo , Viscosidade
17.
Electrophoresis ; 40(10): 1457-1477, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30676660

RESUMO

Circulating tumor cells (CTCs) play an essential role in the metastasis of tumors, and thus can serve as a valuable prognostic factor for malignant diseases. As a result, the ability to isolate and characterize CTCs is essential. This review underlines the potential of dielectrophoresis for CTCs enrichment. It begins by summarizing the key performance parameters and challenges of CTCs isolation using microfluidics. The two main categories of CTCs enrichment-affinity-based and label-free methods-are analysed, emphasising the advantages and disadvantages of each as well as their clinical potential. While the main argument in favour of affinity-based methods is the strong specificity of CTCs isolation, the major advantage of the label-free technologies is in preserving the integrity of the cellular membrane, an essential requirement for downstream characterization. Moving forward, we try to answer the main question: "What makes dielectrophoresis a method of choice in CTCs isolation?" The uniqueness of dielectrophoretic CTCs enrichment resides in coupling the specificity of the isolation process with the conservation of the membrane surface. The specificity of the dielectrophoretic method stems from the differences in the dielectric properties between CTCs and other cells in the blood: the capacitances of the malignantly transformed cellular membranes of CTCs differ from those of other cells. Examples of dielectrophoretic devices are described and their performance evaluated. Critical requirements for using dielectrophoresis to isolate CTCs are highlighted. Finally, we consider that DEP has the potential of becoming a cytometric method for large-scale sorting and characterization of cells.


Assuntos
Separação Celular/métodos , Eletroforese/métodos , Células Neoplásicas Circulantes/patologia , Células Sanguíneas/citologia , Células Sanguíneas/patologia , Separação Celular/instrumentação , Sobrevivência Celular , Eletrodos , Eletroforese/instrumentação , Desenho de Equipamento , Humanos
18.
J Agric Food Chem ; 67(5): 1402-1408, 2019 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-30629411

RESUMO

Ginsenoside compound K (CK) is not a ginsenoside that naturally exists in Panax ginseng Meyer. However, CK is a major metabolite of ginsenoside Rb1, Rb2, or Rc in the intestine under the effects of bacteria. In this study, we first investigated the effects of CK on myelosuppression in mice induced by cyclophosphamide (CTX). The respective quantities of white blood cells, blood platelets, and bone marrow nucleated cells (BMNCs) were determined to be 8.54 ± 0.91 (109/L), 850.90 ± 44.11 (109/L), and 1.45 ± 0.22 (109/L) in the CK-H group by detecting peripheral blood cells and BMNCs. CK-H and CK-L both increased the thymus index by up to 0.62 ± 0.06 (mg/g) and 0.52 ± 0.09 (mg/g), respectively, and significantly increased the yields of colony formation units-granulocyte monocyte and colony formation units-megakaryocytic. According to our study, CK could control apoptosis and promote cells to enter the normal cell cycle by the bcl-2/bax signaling pathway and MEK/ERK signaling pathway. Therefore, the BMNCs could proliferate and differentiate normally after entering the normal cell cycle. So the peripheral blood cells could show a trend of returning to normal. The recovery of peripheral blood cells resulting in the level of cytokines tended to normal. This process may be the mechanisms of CK on myelosuppression. This study provides a reference for ginseng in the treatment of myelosuppression.


Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Ginsenosídeos/farmacologia , Células Mieloides/efeitos dos fármacos , Mielopoese/efeitos dos fármacos , Panax/química , Animais , Apoptose/efeitos dos fármacos , Células Sanguíneas/citologia , Células Sanguíneas/efeitos dos fármacos , Ciclo Celular , Células Cultivadas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Células Mieloides/citologia , Baço/citologia , Baço/efeitos dos fármacos , Timo/citologia , Timo/efeitos dos fármacos
19.
Methods Mol Biol ; 1930: 11-17, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30610593

RESUMO

Peripheral blood is the most common source of T-lymphocytes for in vitro culture. Here, we present a simple and standardized method for small- or large-scale isolation of viable T-lymphocytes and other mononuclear cells from fresh peripheral blood or buffy coat blood samples using the density gradient centrifugation. T-cells obtained using the protocol described here can be used for a variety of downstream analysis, including cellular, molecular, and functional assays.


Assuntos
Células Sanguíneas/citologia , Separação Celular/métodos , Centrifugação com Gradiente de Concentração/métodos , Leucócitos Mononucleares/citologia , Linfócitos T/citologia , Ficoll , Humanos
20.
In Vitro Cell Dev Biol Anim ; 55(2): 104-112, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30617572

RESUMO

Human menstrual blood-derived mesenchymal stem cells (MenSCs) hold great promise for regenerative medicine. Here, H2O2-associated damage in H9c2 cells was employed as an in vitro ischemia-reperfusion model, and the transwell system was used to explore the beneficial effects of MenSCs on the H2O2-induced damage of myocardial H9c2 cells. H2O2 treatment resulted in decreased viability and migration rate, with increased apoptosis levels in cells. By contrast, upon co-culture with MenSCs, H9c2 cell viability and migration were increased, whereas the apoptotic rate decreased. Additionally, western blot and qRT-PCR showed that MenSCs mediated the anti-apoptotic role by downregulating the pro-apoptotic genes Bax and caspase-3, while upregulating the anti-apoptotic effector Bcl-2. Furthermore, co-culture with MenSCs resulted in elevated expression of N-cadherin after H2O2 treatment. These findings indicate that MenSCs protect H9c2 cells against H2O2-associated programmed cell death and would help develop therapeutic tools for cardiomyocyte apoptosis associated with oxidative stress.


Assuntos
Apoptose/efeitos dos fármacos , Células Sanguíneas/citologia , Citoproteção/efeitos dos fármacos , Peróxido de Hidrogênio/toxicidade , Menstruação/sangue , Células-Tronco Mesenquimais/metabolismo , Animais , Apoptose/genética , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Separação Celular , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Ratos
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