scholarly journals Endothelial cells induce proliferation and bone morphogenic protein mediated differentiation of PDGFRα+ human oligodendrocyte precursor cells to astrocytes in trans-well co-culture

2021 ◽  
Author(s):  
Asmita Dasgupta
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Precursor Cells ◽  
Bmp Signaling ◽  
Demyelinating Diseases ◽  
Oligodendrocyte Precursor Cells ◽  
Human Endothelial Cells ◽  
Humoral Factors ◽  
Astrocytic Differentiation ◽  
Oligodendrocyte Precursor

Abstract Background CD140a /PDGFRα + human oligodendrocyte precursor cells (OPCs) are a lineage of OPCs with proven potential for use in cell therapy against demyelinating diseases. However, little is known about the contribution of human endothelial cells in the biology of PDGFRα + human OPCs in the stem cell niche. Methods Transwell co-culture technique with human umbilical vein endothelial cells (HUVECs) was adopted under proliferative or differentiating conditions to understand the role of endothelial cells in these processes within OPCs. Proliferation was followed by measuring OPC sphere size, count, sphere dissociation followed by cell count and 3H-methyl thymidine incorporation. Differentiation was followed by immunocytochemistry. Taqman gene expression assay for selective soluble factors was performed for the two co-culture partner cells to determine the expression of these factors on the biology of the OPCs in presence of the endothelial cells. Results In co-culture with HUVECs, under proliferative conditions, OPCs show increased proliferation and sphere formation. In contrast, under differentiating conditions, OPCs show increased differentiation to astrocytes, with a concomitant decrease in differentiation to oligodendrocytes, compared to no co-culture controls. Transcript assay for selected humoral factors in the OPCs and HUVECs revealed bone morphogenic proteins (BMPs), endothelin1, growth arrest specific 6 (GAS6), and interleukin 6 (IL6) to be in higher abundance in HUVECs than OPCs. Whereas the OPCs show higher expression for pleiotrophin (PTN), fibroblast growth factor 9 (FGF9), ciliary neurotrophic factor (CNTF), and leukemia inhibitory factor (LIF) compared to the endothelial cells. Among the transcripts analyzed, BMP4 transcripts were the highest in relative abundance in the endothelial cells indicating possibilities of BMPs being the critical mediator of endothelial cell-mediated effects. In agreement to this, Noggin effectively attenuated HUVEC mediated astrocytic differentiation of CD140a /PDGFRα + fetal human OPCs. Conclusion Based on the above results, the study concludes that human endothelial cells can significantly alter the biology of PDGFαR + fetal human OPCs mediated by humoral factors to induce increased proliferation and BMP mediated astrocytic differentiation. It can be secondarily inferred from these conclusions that using pharmacological inhibitors of BMP signaling along with the PDGFRα + fetal OPC transplantation may make these cells more effective in remyelination therapy.

scholarly journals Transplanted miR-219-overexpressing oligodendrocyte precursor cells promoted remyelination and improved functional recovery in a chronic demyelinated model

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Hong-Bin Fan ◽  
Li-Xia Chen ◽  
Xue-Bin Qu ◽  
Chuan-Lu Ren ◽  
Xiu-Xiang Wu ◽  
...  
Keyword(s):  
Cognitive Function ◽  
Embryonic Stem ◽  
Precursor Cells ◽  
Demyelinating Diseases ◽  
Specific Cell ◽  
Oligodendrocyte Precursor Cells ◽  
Oligodendrocyte Differentiation ◽  
Ultrastructural Studies ◽  
Multiple Differentiation ◽  
Oligodendrocyte Precursor

Abstract Oligodendrocyte precursor cells (OPCs) have the ability to repair demyelinated lesions by maturing into myelin-producing oligodendrocytes. Recent evidence suggests that miR-219 helps regulate the differentiation of OPCs into oligodendrocytes. We performed oligodendrocyte differentiation studies using miR-219-overexpressing mouse embryonic stem cells (miR219-mESCs). The self-renewal and multiple differentiation properties of miR219-mESCs were analyzed by the expression of the stage-specific cell markers Nanog, Oct4, nestin, musashi1, GFAP, Tuj1 and O4. MiR-219 accelerated the differentiation of mESC-derived neural precursor cells (NPCs) into OPCs. We further transplanted OPCs derived from miR219-mESCs (miR219-OPCs) into cuprizone-induced chronically demyelinated mice to observe remyelination, which resulted in well-contained oligodendrocyte grafts that migrated along the corpus callosum and matured to express myelin basic protein (MBP). Ultrastructural studies further confirmed the presence of new myelin sheaths. Improved cognitive function in these mice was confirmed by behavioral tests. Importantly, the transplanted miR219-OPCs induced the proliferation of endogenous NPCs. In conclusion, these data demonstrate that miR-219 rapidly transforms mESCs into oligodendrocyte lineage cells and that the transplantation of miR219-OPCs not only promotes remyelination and improves cognitive function but also enhances the proliferation of host endogenous NPCs following chronic demyelination. These results support the potential of a therapeutic role for miR-219 in demyelinating diseases.

Single-Chain and Two-Chain Tissue-Type Plasminogen Activator (t-PA) Bind Differently to Cultured Human Endothelial Cells

1989 ◽  
Vol 62 (02) ◽  
pp. 699-703 ◽  
Author(s):  
Rob J Aerts ◽  
Karin Gillis ◽  
Hans Pannekoek
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Binding Sites ◽  
Umbilical Vein ◽  
Tissue Type ◽  
Single Chain ◽  
Human Endothelial Cells ◽  
Tissue Type Plasminogen Activator ◽  
Type Plasminogen Activator ◽  
Umbilical Vein Endothelial Cells

SummaryIt has recently been shown that the fibrinolytic components plasminogen and tissue-type plasminogen activator (t-PA) both bind to cultured human umbilical vein endothelial cells (HUVEC). After cleavage of t-PA by plasmin, “single-chain” t-PA (sct-PA) is converted into “two-chain” t-PA (tct-PA), which differs from the former in a number of respects. We compared binding of sct-PA and tct-PA to the surface of HUVEC. Removal of t-PA bound to HUVEC by a mild treatment with acid and a subsequent quantification of eluted t-PA both by activity- and immunoradiometric assays revealed that, at concentrations between 10 and 500 nM, HUVEC bind about 3-4 times more sct-PA than tct-PA. At these concentrations, both sct-PA and tct-PA remain active when bound to HUVEC. Mutual competition experiments showed that sct-PA and tct-PA can virtually fully inhibit binding of each other to HUVEC, but that an about twofold higher concentration of tct-PA is required to prevent halfmaximal binding of sct-PA than visa versa. These results demonstrate that sct-PA and tct-PA bind with different affinities to the same binding sites on HUVEC.

Assembly and Activation of the Intrinsic Fibrinolytic Pathway on the Surface of Human Endothelial Cells in Culture

1995 ◽  
Vol 74 (02) ◽  
pp. 698-703 ◽  
Author(s):  
Catherine Lenich ◽  
Ralph Pannell ◽  
Victor Gurewich
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Factor Xii ◽  
Plasminogen Activation ◽  
High Molecular Weight Kininogen ◽  
Human Endothelial Cells ◽  
Intrinsic Pathway ◽  
Local Fibrinolysis ◽  
Umbilical Vein Endothelial Cells ◽  
And Function

SummaryFactor XII has long been implicated in the intrinsic pathway of fibrinolysis, but the mechanism by which it triggers plasminogen activation and targets fibrinolysis has not been established. In the present study, the assembly and function of activated Factor XII (F.XIIa), prourokinase (pro-u-PA), high molecular weight kininogen (H-kininogen), and prekallikrein on human umbilical vein endothelial cells (HUVEC) was investigated. 125I-prekallikrein was shown to bind to HUVEC via receptor-bound H-kininogen in the presence of 50 μM ZnCl2. After the addition of F.XIIa, 78% of the 125I-prekallikrein initially bound to HUVEC was converted to 125I-kallikrein. However, only 6% of the HUVEC-bound 125I-pro-u-PA was thereby activated. This discrepancy was shown to be related to rapid dissociation (>50% within 15 min) of prekallikrein/kallikrein, but not pro-u-PA, from HUVEC. Increasing the level of cell-bound kallikrein increased the portion of cell-bound pro-u-PA activated, indicating that their co-localization was important for this pathway. Finally, F.XIIa was shown to trigger plasminogen activation on HUVEC via this pathway. This assembly of reactants on the endothelium suggests a mechanism whereby local fibrinolysis may be triggered by blood coagulation.

Platelets Stimulate Thromboplastin Synthesis in Human Endothelial Cells

1983 ◽  
Vol 49 (02) ◽  
pp. 069-072 ◽  
Author(s):  
U L H Johnsen ◽  
T Lyberg ◽  
K S Galdal ◽  
H Prydz
Keyword(s):  
Endothelial Cells ◽  
De Novo ◽  
Umbilical Vein ◽  
Time Dependent ◽  
De Novo Synthesis ◽  
Human Endothelial Cells ◽  
Tumor Promotor ◽  
Coagulation Assay ◽  
Platelet Aggregates ◽  
Umbilical Vein Endothelial Cells

SummaryHuman umbilical vein endothelial cells in culture synthesize thromboplastin upon stimulation with phytohaemagglutinin (PHA) or the tumor promotor 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The thromboplastin activity is further strongly enhanced in a time dependent reaction by the presence of gel-filtered platelets or platelet aggregates. This effect was demonstrable at platelet concentrations lower than those normally found in plasma, it may thus be of pathophysiological relevance. The thromboplastin activity increased with increasing number of platelets added. Cycloheximide inhibited the increase, suggesting that de novo synthesis of the protein component of thromboplastin, apoprotein III, is necessary.When care was taken to remove monocytes no thromboplastin activity and no apoprotein HI antigen could be demonstrated in suspensions of gel-filtered platelets, platelets aggregated with thrombin or homogenized platelets when studied with a coagulation assay and an antibody neutralization technique.

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