The role of platelet α-granular proteins in the regulation of thrombopoietin messenger RNA expression in human bone marrow stromal cells

Blood ◽  
2000 ◽  
Vol 95 (10) ◽  
pp. 3094-3101 ◽  
Author(s):  
Ranita Sungaran ◽  
Orin T. Chisholm ◽  
Boban Markovic ◽  
Levon M. Khachigian ◽  
Yoshihiro Tanaka ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Mrna Expression ◽  
Stromal Cells ◽  
Messenger Rna ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Dependent Manner ◽  
Platelet Factor ◽  
Dose Dependent

Thrombopoietin (TPO), the specific cytokine that regulates platelet production, is expressed in human bone marrow (BM), kidney, and liver. There appears to be no regulation of TPO in the kidney and liver, but TPO messenger RNA (mRNA) expression can be modulated in the stromal cells of the BM. In this study, we used primary human BM stromal cells as a model to study the regulation of TPO mRNA expression in response to various platelet -granular proteins. We showed that platelet-derived growth factor (PDGF) BB and fibroblast growth factor (FGF) 2 stimulated TPO mRNA expression in both a dose-dependent and time-dependent manner. The addition of 50 ng/mL of PDGF and 20 ng/mL of FGF resulted in maximal induction of TPO mRNA expression in 4 hours. We also found that platelet factor 4 (PF4), thrombospondin (TSP), and transforming growth factor-beta (TGF-β) are negative modulators of megakaryocytopoiesis. We observed suppression in TPO mRNA expression with 1 μg/mL of both PF4 and TSP and 50 ng/mL of TGF-β, with maximal suppression occurring 4 hours after the addition of these proteins. Finally, the addition of whole-platelet lysate produced a dose-dependent inhibition of TPO expression. On the basis of these findings, we propose that the platelet -granular proteins studied may regulate TPO gene expression in BM stromal cells by means of a feedback mechanism.

The role of platelet α-granular proteins in the regulation of thrombopoietin messenger RNA expression in human bone marrow stromal cells

Blood ◽  
2000 ◽  
Vol 95 (10) ◽  
pp. 3094-3101 ◽  
Author(s):  
Ranita Sungaran ◽  
Orin T. Chisholm ◽  
Boban Markovic ◽  
Levon M. Khachigian ◽  
Yoshihiro Tanaka ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Mrna Expression ◽  
Stromal Cells ◽  
Messenger Rna ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Dependent Manner ◽  
Platelet Factor ◽  
Dose Dependent

Abstract Thrombopoietin (TPO), the specific cytokine that regulates platelet production, is expressed in human bone marrow (BM), kidney, and liver. There appears to be no regulation of TPO in the kidney and liver, but TPO messenger RNA (mRNA) expression can be modulated in the stromal cells of the BM. In this study, we used primary human BM stromal cells as a model to study the regulation of TPO mRNA expression in response to various platelet -granular proteins. We showed that platelet-derived growth factor (PDGF) BB and fibroblast growth factor (FGF) 2 stimulated TPO mRNA expression in both a dose-dependent and time-dependent manner. The addition of 50 ng/mL of PDGF and 20 ng/mL of FGF resulted in maximal induction of TPO mRNA expression in 4 hours. We also found that platelet factor 4 (PF4), thrombospondin (TSP), and transforming growth factor-beta (TGF-β) are negative modulators of megakaryocytopoiesis. We observed suppression in TPO mRNA expression with 1 μg/mL of both PF4 and TSP and 50 ng/mL of TGF-β, with maximal suppression occurring 4 hours after the addition of these proteins. Finally, the addition of whole-platelet lysate produced a dose-dependent inhibition of TPO expression. On the basis of these findings, we propose that the platelet -granular proteins studied may regulate TPO gene expression in BM stromal cells by means of a feedback mechanism.

scholarly journals α-Lipoic acid suppresses osteoclastogenesis despite increasing the receptor activator of nuclear factor κB ligand/osteoprotegerin ratio in human bone marrow stromal cells

2005 ◽  
Vol 185 (3) ◽  
pp. 401-413 ◽  
Author(s):  
Jung-Min Koh ◽  
Young-Sun Lee ◽  
Chang-Hyun Byun ◽  
Eun-Ju Chang ◽  
Hyunsoo Kim ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Lipoic Acid ◽  
Nuclear Factor Κb ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Mouse Bone Marrow ◽  
Dependent Manner ◽  
Nuclear Factor ◽  
Receptor Activator ◽  
Dose Dependent

Growing evidence has shown a biochemical link between increased oxidative stress and reduced bone density. Although α-lipoic acid (α-LA) has been shown to act as a thiol antioxidant, its effect on bone cells has not been determined. Using proteomic analysis, we identified six differentially expressed proteins in the conditioned media of α-LA-treated human bone marrow stromal cell line (HS-5). One of these proteins, receptor activator of nuclear factor κB ligand (RANKL), was significantly up-regulated, as confirmed by immunoblotting with anti-RANKL antibody. ELISA showed that α-LA stimulated RANKL production in cellular extracts (membranous RANKL) about 5-fold and in conditioned medium (soluble RANKL) about 23-fold, but had no effect on osteoprotegerin (OPG) secretion. Despite increasing the RANKL/OPG ratio, α-LA showed a dose-dependent suppression of osteoclastogenesis, both in a coculture system of mouse bone marrow cells and osteoblasts and in a mouse bone marrow cell culture system, and reduced bone resorption in a dose-dependent manner. In addition, α-LA-induced soluble RANKL was not inhibited by matrix metalloprotease inhibitors, indicating that soluble RANKL is produced by α-LA without any posttranslational processing. In contrast, α-LA had no significant effect on the proliferation and differentiation of HS-5 cells. These results suggest that α-LA suppresses osteoclastogenesis by directly inhibiting RANKL–RANK mediated signals, not by mediating cellular RANKL production. In addition, our findings indicate that α-LA-induced soluble RANKL is not produced by shedding of membranous RANKL.

scholarly journals Remodeling of Human Osteochondral Defects via rAAV-Mediated Co-Overexpression of TGF-β and IGF-I from Implanted Human Bone Marrow-Derived Mesenchymal Stromal Cells

2019 ◽  
Vol 8 (9) ◽  
pp. 1326
Author(s):  
Stephanie Morscheid ◽  
Jagadeesh Kumar Venkatesan ◽  
Ana Rey-Rico ◽  
Gertrud Schmitt ◽  
Magali Cucchiarini
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Gene Transfer ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Human Model ◽  
Cartilage Lesions ◽  
Igf I

The application of chondrogenic gene sequences to human bone marrow-derived mesenchymal stromal cells (hMSCs) is an attractive strategy to activate the reparative activities of these cells as a means to enhance the processes of cartilage repair using indirect cell transplantation procedures that may improve the repopulation of cartilage lesions. In the present study, we examined the feasibility of co-delivering the highly competent transforming growth factor beta (TGF-β) with the insulin-like growth factor I (IGF-I) in hMSCs via recombinant adeno-associated virus (rAAV) vector-mediated gene transfer prior to implantation in a human model of osteochondral defect (OCD) ex vivo that provides a microenvironment similar to that of focal cartilage lesions. The successful co-overexpression of rAAV TGF-β/IGF-I in implanted hMSCs promoted the durable remodeling of tissue injury in human OCDs over a prolonged period of time (21 days) relative to individual gene transfer and the control (reporter lacZ gene) treatment, with enhanced levels of cell proliferation and matrix deposition (proteoglycans, type-II collagen) both in the lesions and at a distance, while hypertrophic, osteogenic, and catabolic processes could be advantageously delayed. These findings demonstrate the value of indirect, progenitor cell-based combined rAAV gene therapy to treat human focal cartilage defects in a natural environment as a basis for future clinical applications.

Growth factor induced fibroblast differentiation from human bone marrow stromal cells in vitro

2005 ◽  
Vol 23 (1) ◽  
pp. 164-174 ◽  
Author(s):  
Jodie E. Moreau ◽  
Jingsong Chen ◽  
Diah S. Bramono ◽  
Vladimir Volloch ◽  
Herman Chernoff ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Marrow Stromal Cells ◽  
Human Bone ◽  
Human Bone Marrow

Insulin-Like Growth Factor 1 Induced Expression of Vascular Endothelial Growth Factor, Which Is Dependent on MAP Kinase and Phosphatidylinositol 3-Kinase Signaling in Human Bone Marrow Stromal Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4305-4305
Author(s):  
Gexiu Liu ◽  
Guangxiao Tang
Keyword(s):  
Bone Marrow ◽  
Growth Factor ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Brain Injuries ◽  
Marrow Stromal Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Vegf Mrna ◽  
Vegf Protein

Abstract INTRODUCTION: Administration of bone marrow stromal cells after traumatic brain injury provides functional benefit. Its protective mechanisms include neurogenesis and angiogenesis. Different traumatic brain injuries have shown that vascular endothelial growth factor (VEGF) induced neuroprotection, neurogenesis, and angiogenesis. Insulin-like growth factor 1 (IGF-1) is a 7.5-kDa peptide with structural homology to proinsulin. In the adult organism, the liver is the major source of IGF-1. Although IGF-1 is very low in the normal adult rat brain, a number of studies have shown that IGF-1 is strongly induced in the CNS, and exerted its mitogenic and trophic effects on a variety of cell-types after different traumatic brain injuries, suggesting its repair roles after brain damage. We tested the hypothesis that IGF-1 induces expression of VEGF in human bone marrow stromal cells, and its signaling pathway. METHODS: Human bone marrow adherent cells were cultured, and were passaged in DMEM/F12 containing 10% FBS. The fifth passage cells were identified as stromal cells. Subconfluent cells were used, and were washed twice in PBS, then cultured in serum-free DMEM/F12. After overnight incubation, cells were exposed to IGF-1 (100 ng/ml) in the presence of no kinase inhibitor or a 1 h pretreatment with 50 μM PD98059, or 200 nM wortmannin. Cells were harvested after 2 h for analysis of active phosphorylated kinases (Akt and MAPK) or after 24 h for analysis of VEGF protein and mRNA. VEGF mRNA was detected by semi-quantitative RT-PCR, and VEGF protein by ELISA, and kinases by Western blot. RESULTS: 100 ng/ml IGF-1 increased significantly both VEGF mRNA and VEGF protein, which were very low level in control cells. 24 hours after treatment, ratio of RT-PCR product between VEGF and β-actin reach to (38.93±6.73)% from (18.61±4.25)% of control cells (P<0.01). VEGF protein increased to (123.45±20.86)pg/ml from (46.97±8.91)pg/ml (P<0.01). Moreover, IGF-1 induced active phosphorylated kinases (Akt and MAPK). And PD98059 or wortmannin inhibited effects of IGF-1. CONCLUSION: IGF-1 enhanced expression of VEGF mRNA and VEGF protein in human bone marrow stromal cells, which is dependent on MAP kinase and phosphatidylinositol 3-kinase signaling. These results suggest that VEGF be involved in therapeutic effects of bone marrow stromal cells in ischemic disorders, including stroke.

Sign in / Sign up

Export Citation Format

Share Document