Stem cell growth factor: in situ hybridization analysis on the gene expression, molecular characterization and in vitro proliferative activity of a recombinant preparation on primitive hematopoietic progenitor cells

We investigated the effects of stem cell factor (SCF) on the migration of murine bone marrow hematopoietic progenitor cells (HPC) in vitro using a modification of the checkerboard assay. Chemotactic and chemokinetic activities of SCF on HPC were evaluated by the numbers of HPC migrated on positive and negative gradients of SCF, respectively. On both positive and negative gradients of SCF, HPC began to migrate after 4 hours incubation, and their numbers then increased time- dependently. These results indicated that SCF functions as a chemotactic and chemokinetic agent for HPC. Analysis of types of colonies derived from the migrated HPC showed that SCF had chemotactic and chemokinetic effects on all types of HPC. When migrating activities of other cytokines were examined, interleukin (IL)-3 and IL-11 also affected the migration of HPC, but the degrees of each effect were lower than that of SCF. The results of the present study demonstrated that SCF is one of the most potent chemotactic and chemokinetic factors for HPC and suggest that SCF may play an important role in the flow of HPC into bone marrow where stromal cells constitutively produce SCF.

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Gene Expression Analysis of Hematopoietic Progenitor Cells Identifies Dlg7as a Potential Stem Cell Gene

Stem Cells ◽

10.1634/stemcells.2005-0479 ◽

2007 ◽

Vol 25 (6) ◽

pp. 1498-1506 ◽

Cited By ~ 24

Author(s):

Kristbjorn Orri Gudmundsson ◽

Leifur Thorsteinsson ◽

Olafur E. Sigurjonsson ◽

Jonathan R. Keller ◽

Karl Olafsson ◽

...

Keyword(s):

Gene Expression ◽

Stem Cell ◽

Progenitor Cells ◽

Expression Analysis ◽

Gene Expression Analysis ◽

Hematopoietic Progenitor Cells ◽

Hematopoietic Progenitor ◽

Cell Gene ◽

Stem Cell Gene

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Gene expression analysis by non-radioactive RNA-RNA in situ hybridization techniques

Jugoslovenska medicinska biohemija ◽

10.2298/jmh0402127d ◽

2004 ◽

Vol 23 (2) ◽

pp. 127-133 ◽

Cited By ~ 1

Author(s):

Danijela Drakulic ◽

Milena Stevanovic ◽

Gordana Nikcevic

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Cultured Cells ◽

Specific Gene ◽

Rna In Situ Hybridization ◽

Sox Gene ◽

Labeled Rna ◽

Set Up

RNA-RNA in situ hybridization is a reliable method for studying tissue and cell specific gene expression, which enables visualization of labeled antisense RNA probe hybridized to specific mRNA. In this study we employed non-radioactive RNA-RNA in situ hybridization using biotin- or digoxigenin-labeled RNA probes in order to detect SOX gene expression in carcinoma cell lines. By this approach we confirmed results obtained by Northern blot analysis, where the presence of SOX2 mRNA in NT2/D1 and SOX14 mRNA in HepG2 cells has been established. Our aim was to set up RNA-RNA in situ hybridization method in in vitro cultured cells in order to perform further analyses of SOX gene expression on various normal and cancer tissues.

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Hepatocyte growth factor is a synergistic factor for the growth of hematopoietic progenitor cells

Blood ◽

10.1182/blood.v80.10.2454.2454 ◽

1992 ◽

Vol 80 (10) ◽

pp. 2454-2457 ◽

Cited By ~ 99

Author(s):

TE Kmiecik ◽

JR Keller ◽

E Rosen ◽

GF Vande Woude

Keyword(s):

Growth Factor ◽

Epithelial Cells ◽

Hepatocyte Growth Factor ◽

Progenitor Cells ◽

Messenger Rna ◽

Hematopoietic Progenitor Cells ◽

Hematopoietic Progenitor Cell ◽

Hematopoietic Progenitor ◽

Hepatocyte Growth

Abstract Bone marrow (BM) stromal cells, which include macrophages, fibroblasts, endothelial cells, and adipocytes, have been shown to produce several factors that modulate the growth of BM progenitors. Hepatocyte growth factor (HGF) is a fibroblast-derived factor and has recently been shown to be a ligand for the c-met proto-oncogene, a member of the receptor class of tyrosine kinases. c-met messenger RNA (mRNA) is predominantly expressed in epithelial cells, but has been detected in several murine hematopoietic progenitor cell lines, suggesting that HGF and met might function during hematopoiesis. Here, BM cells were found to express both met mRNA and protein. Moreover, HGF was shown to synergize with interleukin-3 and granulocyte-macrophage colony-stimulating factor to stimulate colony formation of hematopoietic progenitor cells in vitro. These results show that, in addition to its activity on epithelial cells, HGF is a new member of the functionally related group of factors that modulate hematopoiesis.

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Abstract 1504: Increased Expression of Stem Cell Factor and its Receptor Following LVAD: A Potential Novel Target for Therapeutic Interventions In Heart Failure

Circulation ◽

10.1161/circ.116.suppl_16.ii_310-b ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Jama Jahanyar ◽

Keith A Youker ◽

George P Noon ◽

Guillermo Torre-Amione ◽

Michael M Koerner ◽

...

Keyword(s):

Gene Expression ◽

Heart Failure ◽

Stem Cell ◽

In Situ Hybridization ◽

Mast Cells ◽

Cardiac Remodeling ◽

Stem Cell Factor ◽

Therapeutic Interventions ◽

Circulatory Support

Introduction: Previous studies have demonstrated the involvement of mast cells (MCs) in cardiac remodeling during heart failure. LVADs cause an influx of MCs into the failing heart, but the underlying mechanism is unknown. We hypothesize that stem cell factor (SCF) induces migration of MCs to the heart. This study investigates the potential role of SCF and its receptor (C-Kit) in promoting the recruitment of stem cell derived MCs during heart failure and following LVAD support. Methods: Myocardial samples were collected from 10 patients undergoing LVAD implantation (Pre-LVAD) paired with samples taken at the time of heart transplantation (Post-LVAD). Biopsies of 4 normal hearts served as controls. We assessed gene expression of SCF and C-Kit. Additionally, we stained for SCF, C-Kit and tryptase protein and utilized In-situ hybridization to determine the origin of SCF. Results: Real-time PCR: SCF mRNA is significantly increased (P<0.01) following mechanical circulatory support as compared to paired heart failure tissue. C-Kit mRNA was significantly increased post-LVAD, as compared to normal tissues (p<0.05). Immunohistochemistry: The C-Kit protein was only expressed on cardiac mast cells. In-Situ hybridization: SCF mRNA was found in endothelial cells, myocytes and interstitial cells. This was confirmed by antibody staining for the SCF protein. Conclusions: LVADs cause an increase of SCF and C-Kit gene expression during unloading. SCF appears to be an important mechanism for the recruitment and maturation of MCs involved in cardiac remodeling, and we suggest that pharmacologic or biologic modification of SCF may provide a new therapeutic path for heart failure treatment.

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Hepatocyte growth factor mobilizes and recruits hematopoietic progenitor cells into liver through a stem cell factor-mediated mechanism

Hepatology Research ◽

10.1111/j.1872-034x.2010.00647.x ◽

2010 ◽

Vol 40 (7) ◽

pp. 711-719 ◽

Cited By ~ 13

Author(s):

Fumihito Tajima ◽

Hiroyuki Tsuchiya ◽

Kenichi Nishikawa ◽

Motoyuki Kataoka ◽

Ichiro Hisatome ◽

...

Keyword(s):

Stem Cell ◽

Growth Factor ◽

Hepatocyte Growth Factor ◽

Progenitor Cells ◽

Stem Cell Factor ◽

Hematopoietic Progenitor Cells ◽

Hematopoietic Progenitor ◽

Hepatocyte Growth

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Connective Tissue Growth Factor Is Up-Regulated in the Diabetic Retina: Amelioration by Angiotensin-Converting Enzyme Inhibition

Endocrinology ◽

10.1210/en.2003-0967 ◽

2004 ◽

Vol 145 (2) ◽

pp. 860-866 ◽

Cited By ~ 48

Author(s):

Christos Tikellis ◽

Mark E. Cooper ◽

Stephen M. Twigg ◽

Wendy C. Burns ◽

Mary Tolcos

Keyword(s):

Gene Expression ◽

In Situ Hybridization ◽

Growth Factor ◽

Connective Tissue ◽

Angiotensin Converting Enzyme ◽

Ace Inhibitor ◽

Tissue Growth ◽

Converting Enzyme ◽

Retinal Pathology

Abstract Connective tissue growth factor (CTGF) has been postulated to have prosclerotic and angiogenic properties. The aim of this present study was to characterize retinal CTGF expression in the absence and presence of diabetes and in the context of treatment with the angiotensin-converting enzyme (ACE) inhibitor, perindopril. Retinas were obtained from control, diabetic, and diabetic plus perindopril-treated (3 mg/d) rats. CTGF gene expression was quantitated by RT-PCR and localized by in situ hybridization. CTGF protein expression was analyzed by Western blotting and localized by immunohistochemistry. Diabetes was associated with a greater than 2-fold increase in CTGF mRNA levels, which was attenuated by perindopril treatment. CTGF immunoreactivity was increased almost 2-fold in diabetes and was ameliorated by the ACE inhibitor perindopril. By in situ hybridization and immunohistochemistry, the major site of CTGF gene expression in the retina of diabetic rats was the ganglion cell layer. Based on the known in vivo effects of CTGF, it is postulated that this growth factor plays a pivotal role in mediating diabetes-associated retinal pathology. Furthermore, the protective effects of ACE inhibitors on retinal pathology may partly be mediated via effects on retinal CTGF expression.

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Analysis of Gene Expression Patterns of Epigenetic Enzymes Dnmt3a, Tet1 and Ogt in Murine Chondrogenic Models

Cells ◽

10.3390/cells10102678 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2678

Author(s):

Judit Vágó ◽

Katalin Kiss ◽

Edina Karanyicz ◽

Roland Takács ◽

Csaba Matta ◽

...

Keyword(s):

Gene Expression ◽

Dna Methylation ◽

In Situ Hybridization ◽

Marker Genes ◽

Specific Gene ◽

Cartilage Formation ◽

Methylation Inhibitor ◽

Dna Methylation Inhibitor

We investigated the gene expression pattern of selected enzymes involved in DNA methylation and the effects of the DNA methylation inhibitor 5-azacytidine during in vitro and in vivo cartilage formation. Based on the data of a PCR array performed on chondrifying BMP2-overexpressing C3H10T1/2 cells, the relative expressions of Tet1 (tet methylcytosine dioxygenase 1), Dnmt3a (DNA methyltransferase 3), and Ogt (O-linked N-acetylglucosamine transferase) were further examined with RT-qPCR in murine cell line-based and primary chondrifying micromass cultures. We found very strong but gradually decreasing expression of Tet1 throughout the entire course of in vitro cartilage differentiation along with strong signals in the cartilaginous embryonic skeleton using specific RNA probes for in situ hybridization on frozen sections of 15-day-old mouse embryos. Dnmt3a and Ogt expressions did not show significant changes with RT-qPCR and gave weak in situ hybridization signals. The DNA methylation inhibitor 5-azacytidine reduced cartilage-specific gene expression and cartilage formation when applied during the early stages of chondrogenesis. In contrast, it had a stimulatory effect when added to differentiated chondrocytes, and quantitative methylation-specific PCR proved that the DNA methylation pattern of key chondrogenic marker genes was altered by the treatment. Our results indicate that the DNA demethylation inducing Tet1 plays a significant role during chondrogenesis, and inhibition of DNA methylation exerts distinct effects in different phases of in vitro cartilage formation.

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