scholarly journals Granulocyte colony-stimulating factor induction of normal human bone marrow progenitors results in neutrophil-specific gene expression

Blood ◽  
1995 ◽  
Vol 85 (3) ◽  
pp. 799-803 ◽  
Author(s):  
N Berliner ◽  
A Hsing ◽  
T Graubert ◽  
F Sigurdsson ◽  
M Zain ◽  
...  
Keyword(s):  
Gene Expression ◽  
Specific Gene ◽  
Specific Gene Expression ◽  
Hla Dr ◽  
Normal Human ◽  
Granule Proteins ◽  
Stimulating Factor ◽  
Secondary Granule ◽  
Normal Human Bone Marrow

We have used a combination of hematopoietic growth factors to induce in vitro granulocytic maturation. A fraction of marrow cells enriched for hematopoietic progenitor cells (CD34+, HLA-DR+) was isolated from normal human bone marrow by monoclonal antibody staining and fluorescence-activated cell sorting. Cells were cultured in a suspension system for 3 days in the presence of stem cell factor and interleukin-3 (IL-3), after which granulocyte colony-stimulating factor (G-CSF) was added. Cells were harvested daily and analyzed for phenotypic maturation by morphologic criteria, and total RNA was obtained for analysis of myeloid gene expression. Maturation was observed to progress to the late metamyelocyte and band stage over a period of 10 to 12 days. Neutrophil-specific gene expression was assayed by reverse transcription-polymerase chain reaction (RT-PCR). Induction with G-CSF resulted in sequential expression of primary and secondary granule proteins, with asynchronous expression of primary granule proteins starting from days 1 to 5, and synchronous expression of lactoferrin and transcobalamin I (secondary granule proteins) from days 7 to 8. Interestingly, myeloperoxidase (MPO) mRNA expression was easily detected in both the freshly isolated CD34+, HLA-DR+ cells and cells at all subsequent stages of induction. This suggests that MPO mRNA is expressed very early during neutrophil development, perhaps before the development of significant numbers of phenotypically recognizable granules. This recapitulation of a program of sequential expression of primary and secondary granule protein genes suggests that in vitro marrow culture suspensions to which appropriate growth factors are added can mimic normal granulocytic maturation. This system should provide an important model for the study of neutrophil-specific gene expression.

scholarly journals Microvesicle Induction of Prostate Specific Gene Expression in Normal Human Bone Marrow Cells

2010 ◽  
Vol 184 (5) ◽  
pp. 2165-2171 ◽  
Author(s):  
Joseph F. Renzulli ◽  
Michael Del Tatto ◽  
Gerri Dooner ◽  
Jason Aliotta ◽  
Lisa Goldstein ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Specific Gene ◽  
Specific Gene Expression ◽  
Normal Human ◽  
Normal Human Bone Marrow ◽  
Marrow Cells

scholarly journals Epigenetic regulation of neural cell differentiation plasticity in the adult mammalian brain

2008 ◽  
Vol 105 (46) ◽  
pp. 18012-18017 ◽  
Author(s):  
Jun Kohyama ◽  
Takuro Kojima ◽  
Eriko Takatsuka ◽  
Toru Yamashita ◽  
Jun Namiki ◽  
...  
Keyword(s):  
Gene Expression ◽  
Target Genes ◽  
Epigenetic Modification ◽  
Mammalian Brain ◽  
Cytokine Signaling ◽  
Specific Gene ◽  
Neural Cells ◽  
Specific Gene Expression

Neural stem/progenitor cells (NSCs/NPCs) give rise to neurons, astrocytes, and oligodendrocytes. It has become apparent that intracellular epigenetic modification including DNA methylation, in concert with extracellular cues such as cytokine signaling, is deeply involved in fate specification of NSCs/NPCs by defining cell-type specific gene expression. However, it is still unclear how differentiated neural cells retain their specific attributes by repressing cellular properties characteristic of other lineages. In previous work we have shown that methyl-CpG binding protein transcriptional repressors (MBDs), which are expressed predominantly in neurons in the central nervous system, inhibit astrocyte-specific gene expression by binding to highly methylated regions of their target genes. Here we report that oligodendrocytes, which do not express MBDs, can transdifferentiate into astrocytes both in vitro (cytokine stimulation) and in vivo (ischemic injury) through the activation of the JAK/STAT signaling pathway. These findings suggest that differentiation plasticity in neural cells is regulated by cell-intrinsic epigenetic mechanisms in collaboration with ambient cell-extrinsic cues.

scholarly journals Alcohol exposure impairs trophoblast survival and alters subtype-specific gene expression in vitro

Placenta ◽  
2016 ◽  
Vol 46 ◽  
pp. 87-91 ◽  
Author(s):  
J.I. Kalisch-Smith ◽  
J.E. Outhwaite ◽  
D.G. Simmons ◽  
M. Pantaleon ◽  
K.M. Moritz
Keyword(s):  
Gene Expression ◽  
Alcohol Exposure ◽  
Specific Gene ◽  
Specific Gene Expression

Characterization of hematopoietic lineage-specific gene expression by ES cell in vitro differentiation induction system

Blood ◽  
2000 ◽  
Vol 95 (3) ◽  
pp. 870-878 ◽  
Author(s):  
Takumi Era ◽  
Toshiaki Takagi ◽  
Tomomi Takahashi ◽  
Jean-Christophe Bories ◽  
Toru Nakano
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Embryonic Stem ◽  
Experimental System ◽  
Specific Gene ◽  
Specific Gene Expression ◽  
Differentiation Induction ◽  
Lineage Specific Gene ◽  
Megakaryocytic Cell

The continuous generation of mature blood cells from hematopoietic progenitor cells requires a highly complex series of molecular events. To examine lineage-specific gene expression during the differentiation process, we developed a novel method combiningLacZ reporter gene analysis with in vitro hematopoietic differentiation induction from mouse embryonic stem cells. For a model system using this method, we chose the erythroid and megakaryocytic differentiation pathways. Although erythroid and megakaryocytic cells possess distinct functional and morphologic features, these 2 lineages originate from bipotential erythro-megakaryocytic progenitors and share common lineage-restricted transcription factors. A portion of the 5′ flanking region of the human glycoprotein IIb (IIb) integrin gene extending from base −598 to base +33 was examined in detail. As reported previously, this region is sufficient for megakaryocyte-specific gene expression. However, previous reports that used human erythro-megakaryocytic cell lines suggested that one or more negative regulatory regions were necessary for megakaryocyte-specific gene expression. Our data clearly showed that an approximately 200-base enhancer region extending from −598 to −400 was sufficient for megakaryocyte-specific gene expression. This experimental system has advantages over those using erythro-megakaryocytic cell lines because it recapitulates normal hematopoietic cell development and differentiation. Furthermore, this system is more efficient than transgenic analysis and can easily examine gene expression with null mutations of specific genes.

Prolonged down regulation of specific gene expression in nucleus pulposus cell mediated by RNA interference in vitro

2006 ◽  
Vol 24 (6) ◽  
pp. 1271-1278 ◽  
Author(s):  
Kenichiro Kakutani ◽  
Kotaro Nishida ◽  
Koki Uno ◽  
Toru Takada ◽  
Takatoshi Shimomura ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Nucleus Pulposus ◽  
Nucleus Pulposus Cell ◽  
Specific Gene ◽  
Down Regulation ◽  
Specific Gene Expression

scholarly journals Recombinant gibbon interleukin-3 acts synergistically with recombinant human G-CSF and GM-CSF in vitro

Blood ◽  
1988 ◽  
Vol 71 (6) ◽  
pp. 1596-1600 ◽  
Author(s):  
RL Paquette ◽  
JY Zhou ◽  
YC Yang ◽  
SC Clark ◽  
HP Koeffler
Keyword(s):  
Colony Formation ◽  
Clinical Utility ◽  
Bone Marrow Cells ◽  
Soft Agar ◽  
Interleukin 3 ◽  
Gm Csf ◽  
Normal Human ◽  
Stimulating Factor ◽  
Normal Human Bone Marrow

Recombinant gibbon interleukin-3 (IL-3) is a multilineage hematopoietic colony-stimulating factor (CSF) that recently was cloned and found to be highly homologous with human IL-3. Gibbon IL-3, as well as human granulocyte-CSF (G-CSF) and human granulocyte-macrophage CSF (GM-CSF), stimulated normal human bone marrow cells to form myeloid colonies in soft agar in a sigmoidal dose-response manner. When IL-3 was added to increasing concentrations of G-CSF or GM-CSF, synergistic colony formation occurred as compared with the effects of each CSF alone. Synergism was also noted when G-CSF was added with GM-CSF and when all the CSFs were added simultaneously. The combination of IL-3 and GM-CSF was less stimulatory than all the other CSF combinations. At day 11 of culture, IL-3 induced granulocyte-macrophage (38%), eosinophil (30%), granulocyte (18%), and macrophage (14%) colony formation. In summary, gibbon IL-3 is a growth factor that can synergize with other CSFs to enhance proliferation of myeloid-committed progenitors, suggesting that combinations of CSFs may have clinical utility in patients with neutropenia of various etiologies.

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