scholarly journals Direct Cell-Cell Contact between Mesenchymal Stem Cells and Endothelial Progenitor Cells Induces a Pericyte-Like Phenotype In Vitro

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Markus Loibl ◽  
Andreas Binder ◽  
Marietta Herrmann ◽  
Fabian Duttenhoefer ◽  
R. Geoff Richards ◽  
...  
Keyword(s):  
Stem Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Cell Contact ◽  
Endothelial Progenitor ◽  
Direct Cell ◽  
Large Bone ◽  
Large Bone Defects ◽  
Cell Cell

Tissue engineering techniques for the regeneration of large bone defects require sufficient vascularisation of the applied constructs to ensure a sufficient supply of oxygen and nutrients. In our previous work, prevascularised 3D scaffolds have been successfully established by coculture of bone marrow derived stem cells (MSCs) and endothelial progenitor cells (EPCs). We identified stabilising pericytes (PCs) as part of newly formed capillary-like structures. In the present study, we report preliminary data on the interactions between MSCs and EPCs, leading to the differentiation of pericyte-like cells. MSCs and EPCs were seeded in transwell cultures, direct cocultures, and single cultures. Cells were cultured for 10 days in IMDM 10% FCS or IMDM 5% FCS 5% platelet lysate medium. Gene expression of PC markers, CD146, NG2,αSMA, and PDGFR-β, was analysed using RT-PCR at days 0, 3, 7, and 10. The upregulation of CD146, NG2, andαSMA in MSCs in direct coculture with EPCs advocates the MSCs’ differentiation towards a pericyte-like phenotype in vitro. These results suggest that pericyte-like cells derive from MSCs and that cell-cell contact with EPCs is an important factor for this differentiation process. These findings emphasise the concept of coculture strategies to promote angiogenesis for cell-based tissue engineered bone grafts.

scholarly journals Factors Secreted by Mesenchymal Stem Cells and Endothelial Progenitor Cells Have Complementary Effects on Angiogenesis In Vitro

2013 ◽  
Vol 22 (4) ◽  
pp. 643-653 ◽  
Author(s):  
Alexandrina Burlacu ◽  
Gabriela Grigorescu ◽  
Ana-Maria Rosca ◽  
Mihai Bogdan Preda ◽  
Maya Simionescu
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Endothelial Progenitor

scholarly journals Conversion of human adipose derived stem cells into endothelial progenitor cells

2017 ◽  
Vol 4 (3-4) ◽  
pp. 217-227
Author(s):  
Van Hong Tran ◽  
Hoa Trong Nguyen ◽  
Phuc Van Pham
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Vascular Tissue ◽  
Cell Phenotype ◽  
Direct Reprogramming ◽  
Endothelial Progenitor ◽  
Vascular Regeneration

Introduction: Endothelial cells (ECs) or endothelial progenitor cells (EPCs) are essential cells for blood vascular regeneration and vascular tissue engineering. However, the source of EPCs are limited. Indeed, these cells only existence with low rate at some tissues such as bone marrow, umbilical cord blood and peripheral blood. This study aimed to produce EPCs from direct reprogramming of adipose tissue-derived mesenchymal stem cells (ADSCs) by ETV2 transfection in vitro. Methods: ADSCs were isolated according to the published works. They were confirmed as mesenchymal stem cells (MSCs) with some characteristics included expression of CD44, CD73, CD90, negative of CD14, CD45, and HLA-DR; in vitro differentiation into adipocytes, and osteoblasts. ETV-2 mRNA was in vitro produced by commercial kit. ETV-2 mRNA molecules were transfected into ADSCs by Fugenes and Lipofectamine agents. These transfected cells were evaluated the expression of EPC properties included expression of CD31, VEGFR-2 in the cell surface by flow cytometry, immunocytochemistry, and in vitro vessel formation in the Matrigel. Results: The results showed that ETV-2 could transform the ADSCs from mesenchymal cell phenotype into endothelial cell phenotype with 10% transfected ADSCs expressing the CD31 in their surface, they also could form the vessel structure in vitro. Conclusion: Although the low efficacy of direct reprogramming, this study gave the new strategy to produce EPCs from the favorite cell sources as ADSCs.

scholarly journals Mesenchymal Stem Cells Attract Endothelial Progenitor Cells via a Positive Feedback Loop between CXCR2 and CXCR4

2019 ◽  
Vol 2019 ◽  
pp. 1-9
Author(s):  
Yingyun Tan ◽  
Linjing Shu ◽  
Peng Xu ◽  
Shi Bai
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Feedback Loop ◽  
Underlying Mechanism ◽  
Endothelial Progenitor ◽  
Molecular Events

Mesenchymal stem cells (MSCs) can attract host endothelial progenitor cells (EPCs) to promote vascularization in tissue-engineered constructs (TECs). Nevertheless, the underlying mechanism remains vague. This study is aimed at investigating the roles of CXCR2 and CXCR4 in the EPC migration towards MSCs. In vitro, Transwell assays were performed to evaluate the migration of EPCs towards MSCs. Antagonists and shRNAs targeting CXCR2, CXCR4, and JAK/STAT3 were applied for the signaling blockade. Western blot and RT-PCR were conducted to analyze the molecular events in EPCs. In vivo, TECs were constructed and subcutaneously implanted into GFP+ transgenic mice. Signaling inhibitors were injected in an orientated manner into TECs. Recruitment of host CD34+ cells was evaluated by immunofluorescence. Eventually, we demonstrated that CXCR2 and CXCR4 were both highly expressed in migrated EPCs and indispensable for MSC-induced EPC migration. CXCR2 and CXCR4 strongly correlated with each other in the way that the expression of CXCR2 and CXCR2-mediated migration depends on the activity of CXCR4 and vice versa. Further studies documented that both of CXCR2 and CXCR4 activated STAT3 signaling, which in turn regulated the expression of CXCR2 and CXCR4, as well as cell migration. In summary, we firstly introduced a reciprocal crosstalk between CXCR2 and CXCR4 in the context of EPC migration. This feedback loop plays critical roles in the migration of EPCs towards MSCs.

scholarly journals Isolation of endothelial progenitor cells from human adipose tissue

2016 ◽  
Vol 3 (05) ◽  
pp. 645-652 ◽  
Author(s):  
Phuc Van Pham ◽  
Ngoc Bich Vu ◽  
Hoa Trong Nguyen ◽  
Ngoc Kim Phan
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Human Adipose Tissue ◽  
Vascular Medicine ◽  
Endothelial Progenitor ◽  
Adipose Tissues ◽  
Surface Marker Expression

Adipose tissue is a rich source of stem cells, especially mesenchymal stem cells (MSCs). This study aimed to identify and isolate endothelial progenitor cells (EPCs) from human adipose tissue. Belly adipose tissues were collected from donors with consent. Stromal vascular fractions (SVFs) were extracted from adipose tissues by enzyme collagenase using commercial kits. SVFs were cultured in MSCCult medium for 24 h to obtain MSCs, then supernatant was collected and cell pellet cultured in EGM-2 medium to obtain adipose tissue EPCs (ADEPCs). ADEPCs were checked for surface marker expression of CD31 and VEGFR2, and for angiogenesis capability in vitro. The results showed that SVFs contained a pool of EPCs with strong angiogenesis potential and that adipose tissue is not only a source for MSCs but also for EPCs. Therefore, ADEPCs may a useful source of EPCs for vascular medicine.

Gene transfer of endothelial NO-synthase restores migratory capacity of endothelial progenitor cells from patients with coronary artery diseases

2007 ◽  
Vol 30 (4) ◽  
pp. 96
Author(s):  
Michael R. Ward ◽  
Qiuwang Zhang ◽  
Duncan J. Stewart ◽  
Michael J.B. Kutryk
Keyword(s):  
Risk Factors ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Fold Increase ◽  
No Synthase ◽  
Endothelial Progenitor ◽  
Migratory Capacity ◽  
Acute Mi ◽  
Cad Risk

Autologous endothelial progenitor cells (EPCs) have been used extensively in the development of cell-based therapy for acute MI. However, EPCs isolated from patients with CAD and/or CAD risk factors have reduced regenerative activity compared to cells from healthy subjects. As in endothelial cells, endothelial NO synthase (eNOS) expression and subsequent NO production are believed to be critical determinants of EPC function. Recently, the ability of EPCs to migrate in vitro in response to chemotactic stimuli has been shown to predict their regenerative capacity in clinical studies. Therefore, we hypothesized that the regenerative function of EPCs from patients with or at high risk for CAD will be enhanced by overexpression of eNOS, as assessed by migratory capacity. Methods: EPCs were isolated from the blood of human subjects with CAD risk factors (>15% Framingham risk score; FRS) (± CAD) by Ficoll gradient separation and differential culture. Following 3 days in culture, cells were transduced using lentivirus vectors containing either eNOS or GFP (sham) at an MOI of 3. The cells were cultured for an additional 5 days before being used in functional assays. Cell migration and chemotaxis in response to VEGF (50 ng/mL) and SDF-1 (100 ng/mL) were assessed using a modified Boyden Chamber assay. Results: Transduction at an MOI of 3 led to a ~90-100-fold increase in eNOS mRNA expression and a 5-6 fold increase in eNOS protein expression, as assessed by qRT-PCR and Western Blotting. Moreover, there was a significant improvement in the migration of EPCs following eNOS transduction compared to sham-transduced EPCs in response to both VEGF (44.3 ± 8.4 vs. 31.1 ± 4.6 cells/high power field; n=10, p < 0.05) and SDF-1 (51.9 ± 11.1 vs. 34.5 ± 3.3 cells/HPF; n=10, p < 0.05). Conclusions: These data show that the reduced migration capacity of EPCs isolated from patients with CAD and/or CAD risk factors can be significantly improved through eNOS overexpression in these cells. Thus, eNOS transduction of autologous EPCs may enhance their ability to restore myocardial perfusion and function following acute MI. We intend to further explore the regenerative potential of eNOS-transduced EPCs using various in vitro and in vivo models.

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