scholarly journals Bradykinin Protects Human Endothelial Progenitor Cells from High-Glucose-Induced Senescence through B2 Receptor-Mediated Activation of the Akt/eNOS Signalling Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Yuehuan Wu ◽  
Cong Fu ◽  
Bing Li ◽  
Chang Liu ◽  
Zhi He ◽  
...  
Keyword(s):  
Cord Blood ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
High Glucose ◽  
Copy Number ◽  
Relative Length ◽  
Healthy Controls ◽  
Mtdna Copy Number ◽  
Endothelial Progenitor ◽  
Cd34 Cells

Background. Circulating endothelial progenitor cells (EPCs) play important roles in vascular repair. However, the mechanisms of high-glucose- (HG-) induced cord blood EPC senescence and the role of B2 receptor (B2R) remain unknown. Methods. Cord blood samples from 26 patients with gestational diabetes mellitus (GDM) and samples from 26 healthy controls were collected. B2R expression on circulating CD34+ cells of cord blood mononuclear cells (CBMCs) was detected using flow cytometry. The plasma concentrations of 8-isoprostaglandin F2α (8-iso-PGF2α) and nitric oxide (NO) were measured. EPCs were treated with HG (40 mM) alone or with bradykinin (BK) (1 nM). The B2R and eNOS small interfering RNAs (siRNAs) and the PI3K antagonist LY294002 were added to block B2R, eNOS, and PI3K separately. To determine the number of senescent cells, senescence-associated β-galactosidase (SA-β-gal) staining was performed. The level of mitochondrial reactive oxygen species (ROS) in EPCs was assessed by Mito-Sox staining. Cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assays. Mitochondrial DNA (mtDNA) copy number and the relative length of telomeres were detected by real time-PCR. The distribution of human telomerase reverse transcriptase (hTERT) in the nucleus, cytosol, and mitochondria of EPCs was detected by immunofluorescence. The expression of B2R, p16, p21, p53, P-Ser473AKT, T-AKT, eNOS, and hTERT was demonstrated by Western blot. Results. B2R expression on circulating CD34+ cells of CBMCs was significantly reduced in patients with GDM compared to healthy controls. Furthermore, B2R expression on circulating CD34+ cells of CBMCs was inversely correlated with plasma 8-iso-PGF2α concentrations and positively correlated with plasma NO levels. BK treatment decreased EPC senescence and ROS generation. Furthermore, BK treatment of HG-exposed cells led to elevated P-Ser473AKT and eNOS protein expression compared with HG treatment alone. BK reduced hTERT translocation in HG-induced senescent EPCs. B2R siRNA, eNOS siRNA, and antagonist of the PI3K signalling pathway blocked the protective effects of BK. Conclusion. BK, acting through PI3K-AKT-eNOS signalling pathways, reduced hTERT translocation, increased the relative length of telomeres while reducing mtDNA copy number, and finally protected against EPC senescence induced by HG.

scholarly journals Expansion, Differentiation and Transplantation of Human Endothelial Progenitor Cells Derived from Umbilical Cord Blood CD34+ Cells

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1123-1123
Author(s):  
Meng Qin ◽  
Qing-Yu Zhang ◽  
Yupo Ma ◽  
Wei Dai ◽  
Yongping Jiang
Keyword(s):  
Cell Therapy ◽  
Cord Blood ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Basal Medium ◽  
Human Umbilical Cord ◽  
Endothelial Progenitor ◽  
Cd34 Cells

Abstract Transplantation of endothelial progenitor cells (EPCs)/endothelial cells (ECs) has been developed as a cell therapy for ischemic diseases and hemophilia A, due to the capacities of these cells to repair vascular damage and produce factor VIII. The strategies and the mechanisms of this type of cell therapy have been extensively investigated in the past decades. However, given the problems associated with host immune responses to grafts in allotransplantation, the difficulties in obtaining adequate numbers of suitable cells for transplantation represent additional obstacles for a successful cell therapy. In the present study, human umbilical cord blood CD34+ mesenchymal cells (MCs), which generate low immune response, have been adopted as the source of endothelial progenitors. Our major goal is to establish a highly efficient approach to stimulate the expansion and subsequent differentiation of human endothelial progenitor cells (EPCs), in order to obtain sufficient quantities of healthy cells for therapeutic use. The CD34+ cells isolated from human umbilical cord blood were cultured for 6 days in a customized basal medium supplemented with a modified cytokine cocktail, including SCF, Flt-3L, TPO, IL-3, and GM-CSF. At the end of proliferation stage, cell number was calculated with Flow Cytometry, and cell morphology was characterized as well. The numbers of CD34+ cells and CD34+/CD133+/VEGFR-2+ early EPCs cells were found to be increased by approximately 108-fold and 41.9-fold, respectively, on day 6. In the differentiation stage, the attached cells were transferred to EGM-2 basal medium supplemented with FBS and additional nutrients, including VEGF, IGF, EGF, and FGF, for another 15 days. Following this two-step culture, the generated adherent cells were found to have proliferated more than 1200 folds over the initial EPCs, and they were identified as mature endothelial cells expressing CD31, vWF and FVIII. Furthermore, we developed a nonobese diabetic, severe combined immunodeficient (NOD/SCID) mouse model with portal sinusoidal endothelium injury. The NOD/SCID mice were treated with the produced EPCs/ECs through hepatic portal vein injection at the dose of 6x10^6 cells. Tissue examination demonstrated that GFP-labeled transplanted cells migrated and integrated into liver structure, where they expressed specific endothelial cell markers. Therefore, through our unique approach, we can provide an excellent source of healthy endothelial progenitors for cell therapy. Disclosures Qin: Biopharmagen corp: Employment. Jiang:Biopharmagen.corp: Employment.

scholarly journals Short-term phytohaemagglutinin-activated mononuclear cells induce endothelial progenitor cells from cord blood CD34+ cells

2001 ◽  
Vol 113 (4) ◽  
pp. 962-969 ◽  
Author(s):  
Hyoung Jin Kang ◽  
Sung Chun Kim ◽  
Young Ju Kim ◽  
Chul Woo Kim ◽  
Joong Gon Kim ◽  
...  
Keyword(s):  
Cord Blood ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Mononuclear Cells ◽  
Short Term ◽  
Endothelial Progenitor ◽  
Cd34 Cells ◽  
Cord Blood Cd34

scholarly journals Ultrasonic microbubble VEGF gene delivery improves angiogenesis of senescent endothelial progenitor cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yi-Nan Lee ◽  
Yih-Jer Wu ◽  
Hsin-I. Lee ◽  
Hsueh-Hsiao Wang ◽  
Chiung-Yin Chang ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Copy Number ◽  
Therapeutic Effects ◽  
Endothelial Progenitor ◽  
Cell Therapies ◽  
Aortic Endothelial Cells ◽  
Angiogenic Effect ◽  
And Migration

AbstractThe therapeutic effects of ultrasonic microbubble transfection (UMT)-based vascular endothelial growth factor 165 (VEGF165) gene delivery on young and senescent endothelial progenitor cells (EPCs) were investigated. By UMT, plasmid DNA (pDNA) can be delivered into both young EPCs and senescent EPCs. In the UMT groups, higher pDNA-derived protein expression was found in senescent EPCs than in young EPCs. Consistent with this finding, a higher intracellular level of pDNA copy number was detected in senescent EPCs, with a peak at the 2-h time point post UMT. Ultrasonic microbubble delivery with or without VEGF improved the angiogenic properties, including the proliferation and/or migration activities, of senescent EPCs. Supernatants from young and senescent EPCs subjected to UMT-mediated VEGF transfection enhanced the proliferation and migration of human aortic endothelial cells (HAECs), and the supernatant of senescent EPCs enhanced proliferation more strongly than the supernatant from young EPCs. In the UMT groups, the stronger enhancing effect of the supernatant from senescent cells on HAEC proliferation was consistent with the higher intracellular VEGF pDNA copy number and level of protein production per cell in the supernatant from senescent cells in comparison to the supernatant from young EPCs. Given that limitations for cell therapies are the inadequate number of transplanted cells and/or insufficient cell angiogenesis, these findings provide a foundation for enhancing the therapeutic angiogenic effect of cell therapy with senescent EPCs in ischaemic cardiovascular diseases.

scholarly journals Very Small Embryonic-Like Stem Cells, Endothelial Progenitor Cells, and Different Monocyte Subsets Are Effectively Mobilized in Acute Lymphoblastic Leukemia Patients after G-CSF Treatment

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
Andrzej Eljaszewicz ◽  
Lukasz Bolkun ◽  
Kamil Grubczak ◽  
Malgorzata Rusak ◽  
Tomasz Wasiluk ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Acute Lymphoblastic Leukemia ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Lymphoblastic Leukemia ◽  
Monocyte Subsets ◽  
Endothelial Progenitor ◽  
Hematopoietic Stem ◽  
Cd34 Cells

Background. Acute lymphoblastic leukemia (ALL) is a malignant disease of lymphoid progenitor cells. ALL chemotherapy is associated with numerous side effects including neutropenia that is routinely prevented by the administration of growth factors such as granulocyte colony-stimulating factor (G-CSF). To date, the effects of G-CSF treatment on the level of mobilization of different stem and progenitor cells in ALL patients subjected to clinically effective chemotherapy have not been fully elucidated. Therefore, in this study we aimed to assess the effect of administration of G-CSF to ALL patients on mobilization of other than hematopoietic stem cell (HSCs) subsets, namely, very small embryonic-like stem cells (VSELs), endothelial progenitor cells (EPCs), and different monocyte subsets. Methods. We used multicolor flow cytometry to quantitate numbers of CD34+ cells, hematopoietic stem cells (HSCs), VSELs, EPCs, and different monocyte subsets in the peripheral blood of ALL patients and normal age-matched blood donors. Results. We showed that ALL patients following chemotherapy, when compared to healthy donors, presented with significantly lower numbers of CD34+ cells, HSCs, VSELs, and CD14+ monocytes, but not EPCs. Moreover, we found that G-CSF administration induced effective mobilization of all the abovementioned progenitor and stem cell subsets with high regenerative and proangiogenic potential. Conclusion. These findings contribute to better understanding the beneficial clinical effect of G-CSF administration in ALL patients following successful chemotherapy.

Abstract 42: In Vivo Differentiation of Epigenetically Reprogrammed Human Endothelial Progenitor Cells into Cardiomyocytes Enhances Functional and Anatomical Postinfarct Myocardial Repair

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Melissa A Thal ◽  
Prasanna Krishnamurthy ◽  
Alexander R Mackie ◽  
Eneda Hoxha ◽  
Erin Lambers ◽  
...  
Keyword(s):  
Heart Disease ◽  
Ischemic Heart Disease ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Ischemic Heart ◽  
Specific Gene ◽  
Myocardial Repair ◽  
Endothelial Progenitor ◽  
Human Cd34 ◽  
Cd34 Cells

Currently, bone marrow derived endothelial progenitor cells (human CD34+ cells, EPC) are being used clinically to improve vascularization in patients with ischemic heart disease. While it is generally accepted that CD34+ cells predominantly work through a paracrine mechanism, there exists no convincing evidence that these cells trans-differentiate into functional cardiomyocytes (CMC). Since ischemic heart disease leads to substantial loss of CMC, improving cardiomyogenic plasticity of an existing autologous cell therapy is of obvious import. EPC and CMC both differentiate from a common mesodermal progenitor however; during EC-specific lineage differentiation, CMC specific genes are epigenetically silenced. We hypothesized that reprogramming of CD34+ cells using small molecules targeting key epigenetic repressive marks may recapitulate their cardiomyogenic potential. Human CD34+ EPCs were treated with inhibitors of histone deacetylases (valproic acid) for 24 hours followed by an additional 24 hours with the DNA methyltransferase inhibitor (5-Azacytidine). This forty-eight hour treatment led to the reactivation of pluripotency associated and CMC specific mRNA expression while EC specific gene expression was maintained. Intra-myocardial transplantation of a sub-therapeutic dose of reprogrammed CD34+ cells in an acute myocardial infarction mouse model showed significant improvement in LV function compared to the same number of control CD34+ cells that are therapeutically equivalent to no treatment at all. This was histologically supported by de novo CMC differentiation. In addition to increased cardiomyogenic plasticity, drug treatment also enhanced the inherent therapeutic capacity of the CD34+ cells as shown by reduced fibrosis, increased capillary density, increased proliferation, increased cell survival and increased secretion of angiogenic factors. Taken together, our results suggest that epigenetically reprogrammed CD34+ cells are “super-CD34+ cells” that have an enhanced paracrine effect, display a more plastic phenotype and improve post-infarct cardiac repair by both neo-cardiomyogenesis and neovascularization.

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