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Sensors (Basel, Switzerland) Sep 2018Bone marrow-derived mesenchymal stem cells (BMSCs) are an important cell resource for stem cell-based therapy, which are generally isolated and enriched by the...
Bone marrow-derived mesenchymal stem cells (BMSCs) are an important cell resource for stem cell-based therapy, which are generally isolated and enriched by the density-gradient method based on cell size and density after collection of tissue samples. Since this method has limitations with regards to purity and repeatability, development of alternative label-free methods for BMSC separation is desired. In the present study, rapid label-free separation and enrichment of BMSCs from a heterogeneous cell mixture with bone marrow-derived promyelocytes was successfully achieved using a dielectrophoresis (DEP) device comprising saw-shaped electrodes. Upon application of an electric field, HL-60 cells as models of promyelocytes aggregated and floated between the saw-shaped electrodes, while UE7T-13 cells as models of BMSCs were effectively captured on the tips of the saw-shaped electrodes. After washing out the HL-60 cells from the device selectively, the purity of the UE7T-13 cells was increased from 33% to 83.5% within 5 min. Although further experiments and optimization are required, these results show the potential of the DEP device as a label-free rapid cell isolation system yielding high purity for rare and precious cells such as BMSCs.
Topics: Bone Marrow Cells; Cell Separation; Electrophoresis; HL-60 Cells; Humans; Mesenchymal Stem Cells; Time Factors
PubMed: 30205546
DOI: 10.3390/s18093007 -
Blood Mar 2021
Topics: Blood Cell Count; Child; Erythrocytes; Female; Granulocyte Precursor Cells; Humans; Leukemia, Myeloid, Acute
PubMed: 33734334
DOI: 10.1182/blood.2020009744 -
Biological Chemistry Oct 2014Angiotensin-converting enzyme (ACE) plays an important role in blood pressure control. ACE also has effects on renal function, reproduction, hematopoiesis, and several... (Review)
Review
Angiotensin-converting enzyme (ACE) plays an important role in blood pressure control. ACE also has effects on renal function, reproduction, hematopoiesis, and several aspects of the immune response. ACE 10/10 mice overexpress ACE in monocytic cells; macrophages from ACE 10/10 mice demonstrate increased polarization toward a proinflammatory phenotype. As a result, ACE 10/10 mice have a highly effective immune response following challenge with melanoma, bacterial infection, or Alzheimer disease. As shown in ACE 10/10 mice, enhanced monocytic function greatly contributes to the ability of the immune response to defend against a wide variety of antigenic and non-antigenic challenges.
Topics: Animals; Granulocyte Precursor Cells; Immunity, Cellular; Mice; Mice, Knockout; Peptidyl-Dipeptidase A
PubMed: 24633750
DOI: 10.1515/hsz-2013-0295 -
Genes Mar 2023Acute promyelocytic leukemia (APL) pathogenesis is based on gene translocations, which are of high importance in the diagnosis of and proper therapy selection for APL....
Acute promyelocytic leukemia (APL) pathogenesis is based on gene translocations, which are of high importance in the diagnosis of and proper therapy selection for APL. However, in some cases acute myeloid leukemia (AML) demonstrates APL-like morphological features such as atypical promyelocytes accumulation. This type of AML is characterized by the involvement of other family members or completely different genes. In the present study, we used conventional karyotyping, FISH and high-throughput sequencing in a group of 271 de novo AML with atypical promyelocytes accumulation. Of those, 255 cases were shown to carry a typical chromosomal translocation t(15;17)(q24;q21) with chimeric gene formation (94.1%). Other -positive cases exhibited cryptic fusion without t(15;17)(q24;q21) (1.8%, = 5) and variant t(5;17)(q35;q21) translocation with chimeric gene formation (1.5%, = 4). However, 7 -negative AMLs with atypical promyelocytes accumulation were also discovered. These cases exhibited and fusions as well as mutations, e.g., insertion and non-recurrent chromosomal aberrations. Our findings demonstrate the genetic diversity of AML with APL-like morphological features, which is of high importance for successful therapy implementation.
Topics: Humans; Granulocyte Precursor Cells; Leukemia, Promyelocytic, Acute; Leukemia, Myeloid, Acute; Translocation, Genetic; Nuclear Proteins
PubMed: 36980947
DOI: 10.3390/genes14030675 -
Cytometry. Part B, Clinical Cytometry 2011The pathological hallmark of myelodysplastic syndromes (MDS) is marrow dysplasia, which represents the basis of the WHO classification of these disorders. This... (Review)
Review
The pathological hallmark of myelodysplastic syndromes (MDS) is marrow dysplasia, which represents the basis of the WHO classification of these disorders. This classification provides clinicians with a useful tool for defining the different subtypes of MDS and determining individual prognosis. The WHO proposal has raised some concern regarding minimal diagnostic criteria particularly in patients with normal karyotype without robust morphological markers of dysplasia (such as ring sideroblasts or excess of blasts). Therefore, there is clearly a need to refine the accuracy to detect marrow dysplasia. Flow cytometry (FCM) immunophenotyping has been proposed as a tool to improve the evaluation of marrow dysplasia. Rationale for the application of FCM in the diagnostic work up of MDS is that immunophenotyping is an accurate method for quantitative and qualitative evaluation of hematopoietic cells and that MDS have been found to have abnormal expression of several cellular antigens. To become clinically applicable, FCM analysis should be based on parameters with sufficient specificity and sensitivity, data should be reproducible between different operators and the results should be easily understood by clinicians. In this report, we reviewed the most relevant progresses in detection of marrow dysplasia by FCM in MDS as defined by WHO criteria.
Topics: Antigens, CD; Bone Marrow; Erythroid Cells; Flow Cytometry; Granulocyte Precursor Cells; Humans; Immunophenotyping; Myelodysplastic Syndromes; Phenotype; World Health Organization
PubMed: 21674774
DOI: 10.1002/cyto.b.20607 -
Journal of Immunology (Baltimore, Md. :... Sep 2015The production of mature eosinophils (Eos) is a tightly orchestrated process with the aim to sustain normal Eos levels in tissues while also maintaining low numbers of...
The production of mature eosinophils (Eos) is a tightly orchestrated process with the aim to sustain normal Eos levels in tissues while also maintaining low numbers of these complex and sensitive cells in the blood. To identify regulators of homeostatic eosinophilopoiesis in mice, we took a global approach to identify genome-wide transcriptome and epigenome changes that occur during homeostasis at critical developmental stages, including Eos-lineage commitment and lineage maturation. Our analyses revealed a markedly greater number of transcriptome alterations associated with Eos maturation (1199 genes) than with Eos-lineage commitment (490 genes), highlighting the greater transcriptional investment necessary for differentiation. Eos-lineage-committed progenitors (EoPs) were noted to express high levels of granule proteins and contain granules with an ultrastructure distinct from that of mature resting Eos. Our analyses also delineated a 976-gene Eos-lineage transcriptome that included a repertoire of 56 transcription factors, many of which have never previously been associated with Eos. EoPs and Eos, but not granulocyte-monocyte progenitors or neutrophils, expressed Helios and Aiolos, members of the Ikaros family of transcription factors, which regulate gene expression via modulation of chromatin structure and DNA accessibility. Epigenetic studies revealed a distinct distribution of active chromatin marks between genes induced with lineage commitment and genes induced with cell maturation during Eos development. In addition, Aiolos and Helios binding sites were significantly enriched in genes expressed by EoPs and Eos with active chromatin, highlighting a potential novel role for Helios and Aiolos in regulating gene expression during Eos development.
Topics: Animals; Binding Sites; Cell Lineage; Cells, Cultured; Chromatin; Cytoplasmic Granules; DNA-Binding Proteins; Eosinophils; Gene Expression Regulation; Granulocyte Precursor Cells; Hematopoiesis; Ikaros Transcription Factor; Mice; Mice, Inbred BALB C; Trans-Activators; Transcription Factors; Transcriptome
PubMed: 26268651
DOI: 10.4049/jimmunol.1500510 -
Life Science Alliance Jan 2021Chromosomal rearrangements of the mixed-lineage leukemia gene are the hallmark of infant acute leukemia. The granulocyte-macrophage progenitor state forms the...
Chromosomal rearrangements of the mixed-lineage leukemia gene are the hallmark of infant acute leukemia. The granulocyte-macrophage progenitor state forms the epigenetic basis for myelomonocytic leukemia stemness and transformation by MLL-type oncoproteins. Previously, it was shown that the establishment of murine myelomonocytic - transformation, but not its maintenance, depends on the transcription factor C/EBPα, suggesting an epigenetic hit-and-run mechanism of MLL-driven oncogenesis. Here, we demonstrate that compound deletion of / almost entirely abrogated the growth and survival of --transformed cells. Rare, slow-growing, and apoptosis-prone --transformed escapees were recovered from compound / deletions. The escapees were uniformly characterized by high expression of the resident gene, suggesting inferior functional compensation of C/EBPα/C/EBPβ deficiency by C/EBPε. Complementation was augmented by ectopic C/EBPβ expression and downstream activation of IGF1 that enhanced growth. gene inactivation was accomplished only in the presence of complementing C/EBPβ, but not in its absence, confirming the dependency of the / double knockouts. Our data show that -transformed myeloid cells are dependent on C/EBPs during the initiation and maintenance of transformation.
Topics: Animals; Apoptosis; CCAAT-Enhancer-Binding Protein-alpha; CCAAT-Enhancer-Binding Protein-beta; CCAAT-Enhancer-Binding Proteins; Cell Proliferation; Cell Survival; Cell Transformation, Neoplastic; Gene Knockout Techniques; Granulocyte Precursor Cells; HEK293 Cells; Humans; Leukemia, Myeloid, Acute; Mice; Myeloid-Lymphoid Leukemia Protein; Oncogene Proteins, Fusion; Signal Transduction; Transfection
PubMed: 33144337
DOI: 10.26508/lsa.202000709 -
Haematologica Aug 2015Acute promyelocytic leukemia is an aggressive malignancy characterized by the accumulation of promyelocytes in the bone marrow. PML/RARA is the primary abnormality...
Acute promyelocytic leukemia is an aggressive malignancy characterized by the accumulation of promyelocytes in the bone marrow. PML/RARA is the primary abnormality implicated in this pathology, but the mechanisms by which this chimeric fusion protein initiates disease are incompletely understood. Identifying PML/RARA targets in vivo is critical for comprehending the road to pathogenesis. Utilizing a novel sorting strategy, we isolated highly purified promyelocyte populations from normal and young preleukemic animals, carried out microarray and methylation profiling analyses, and compared the results from the two groups of animals. Surprisingly, in the absence of secondary lesions, PML/RARA had an overall limited impact on both the transcriptome and methylome. Of interest, we did identify down-regulation of secondary and tertiary granule genes as the first step engaging the myeloid maturation block. Although initially not sufficient to arrest terminal granulopoiesis in vivo, such alterations set the stage for the later, complete differentiation block seen in leukemia. Further, gene set enrichment analysis revealed that PML/RARA promyelocytes exhibit a subtle increase in expression of cell cycle genes, and we show that this leads to both increased proliferation of these cells and expansion of the promyelocyte compartment. Importantly, this proliferation signature was absent from the poorly leukemogenic p50/RARA fusion model, implying a critical role for PML in the altered cell-cycle kinetics and ability to initiate leukemia. Thus, our findings challenge the predominant model in the field and we propose that PML/RARA initiates leukemia by subtly shifting cell fate decisions within the promyelocyte compartment.
Topics: Animals; Antigens, CD34; Cell Proliferation; Cell Transformation, Neoplastic; Cluster Analysis; DNA Methylation; Disease Models, Animal; Gene Expression Profiling; Gene Expression Regulation, Leukemic; Granulocyte Precursor Cells; Humans; Immunophenotyping; Leukemia, Promyelocytic, Acute; Mice; Mice, Transgenic; Neoplastic Stem Cells; Oncogene Proteins, Fusion; Transcription, Genetic
PubMed: 26088929
DOI: 10.3324/haematol.2014.123018 -
Molecules (Basel, Switzerland) Jan 2022Kaempferol is a well-known antioxidant found in many plants and plant-based foods. In plants, kaempferol is present mainly in the form of glycoside derivatives. In this...
Kaempferol is a well-known antioxidant found in many plants and plant-based foods. In plants, kaempferol is present mainly in the form of glycoside derivatives. In this work, we focused on determining the effect of kaempferol and its glycoside derivatives on the expression level of genes related to the reduction of oxidative stress-, , , , and ; the enzymatic activity of superoxide dismutases; and the level of glutathione. We used HL-60 acute promyelocytic leukemia cells, which were incubated with the anticancer drug etoposide and kaempferol or one of its three glycoside derivatives isolated from the aerial parts of Medik.-kaempferol 3--[(6--caffeoyl)-β-d-glucopyranosyl-(1→2)]-β-d-galactopyranoside-7--β-d-glucuropyranoside (P2), kaempferol 3--[(6--coumaroyl)-β-d-glucopyranosyl-(1→2)]-β-d-galactopyranoside-7--β-d-glucuropyranoside (P5), and kaempferol 3--[(6--feruloyl)-β-d-glucopyranosyl-(1→2)]-β-d-galactopyranoside-7--β-d-glucuropyranoside (P7). We showed that none of the tested compounds affected gene expression. Co-incubation with etoposide (1 µM) and kaempferol (10 and 50 µg/mL) leads to an increase in the expression of the (9.49 and 9.33-fold at 10 µg/mL and 50 µg/mL, respectively), (1.68-fold at 10 µg/mL), (1.72-fold at 10-50 µg/mL), and (1.84-fold at 50 µg/mL) genes in comparison to cells treated only with etoposide. The effect of kaempferol derivatives on gene expression differs depending on the derivative. All tested polyphenols increased the SOD activity in cells co-incubated with etoposide. We observed that the co-incubation of HL-60 cells with etoposide and kaempferol or derivative P7 increases the level of total glutathione in these cells. Taken together, our observations suggest that the antioxidant activity of kaempferol is related to the activation of antioxidant genes and proteins. Moreover, we observed that glycoside derivatives can have a different effect on the antioxidant cellular systems than kaempferol.
Topics: Antineoplastic Agents, Phytogenic; Antioxidants; Drug Therapy, Combination; Etoposide; Glycosides; HL-60 Cells; Humans; Kaempferols; Lens Plant; Leukemia, Promyelocytic, Acute; Oxidative Stress; Plant Extracts
PubMed: 35056649
DOI: 10.3390/molecules27020333 -
Cellular Signalling May 2020The promyelocytic leukemia-retinoic acid receptor α (PML/RARα) is hypothesized to play a vital role in the pathogenesis of acute promyelocytic leukemia (APL). A...
The promyelocytic leukemia-retinoic acid receptor α (PML/RARα) is hypothesized to play a vital role in the pathogenesis of acute promyelocytic leukemia (APL). A previous study has demonstrated that PML/RARα is cleaved by neutrophil elastase (NE) in early myeloid cells, which leads to an increase in the nuclear localization signal (NLS) in RARα and in the incidence of APL. In this study, we explored the effects of NLS-RARα on acute myeloid leukemia (AML) cells and studied the mechanism of its localization. LV-NLS-RARα recombinant lentivirus and negative control LV-NC lentivirus were transfected into HL-60 cells and U937 cells while mutant NLS-RARα were transfected into U937 cells, and all groups were treated with 1α, 25-dihydroxyvitamin D3(1,25D3). The results showed that NLS-RARα was located mainly in the nucleus while mutant NLS-RARα was located in the cytoplasm. Overexpression of NLS-RARα downregulated the expression of CD11b, CD11c, CD14, and three forms of CEBPβ compared to the overexpression of NC and mutant NLS-RARα. It was speculated that the abnormal localization of NLS-RARα was mediated via importin-α/β in the pathogenesis of APL. By producing point mutations in the two NLSs in NLS-RARα, we showed that the nuclear import of NLS-RARα was mainly dependent on the NLS of the RARα portion. Subsequently, we found that importin-α1 (KPNA2)/importin-β1 (KPNB1) participates in the nuclear transport of NLS-RARα. Taken together, abnormal localization of NLS-RARα blocks the differentiation of APL cells, and nuclear localization of NLS-RARα depends on NLS of the RARα portion and is mediated via binding with importin-α/β.
Topics: Active Transport, Cell Nucleus; Cell Nucleus; HL-60 Cells; Humans; Leukemia, Promyelocytic, Acute; Nuclear Localization Signals; Retinoic Acid Receptor alpha; U937 Cells; alpha Karyopherins; beta Karyopherins
PubMed: 32036017
DOI: 10.1016/j.cellsig.2020.109567