Endothelial colony forming cells generated from cryopreserved peripheral blood mononuclear cells

Béland, Stéphanie; Désy, Olivier; Bouchard-Boivin, François; Gama, Alcino; Serres, Sacha A. De

doi:10.1016/j.humimm.2021.02.009

Cited by 3 publications

(4 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is noteworthy that we have used for this study cryopreserved EPCs obtained from freshly isolated PBMCs. Despite the fact that the effect of cryopreservation on EPCs’ physiology is still controversial [ 68 – 70 ], such strategy allows the creation of a bank of EPCs from different donors or sources (e.g. bone marrow, blood, umbilical cord) for screening the cells with better angiogenic activity.…”

Section: Discussionmentioning

confidence: 99%

RETRACTED ARTICLE: Secretome of endothelial progenitor cells from stroke patients promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage

et al. 2021

View full text Add to dashboard Cite

Background Cell-based therapeutic strategies have been proposed as an alternative for brain repair after stroke, but their clinical application has been hampered by potential adverse effects in the long term. The present study was designed to test the effect of the secretome of endothelial progenitor cells (EPCs) from stroke patients (scCM) on in vitro human models of angiogenesis and vascular barrier. Methods Two different scCM batches were analysed by mass spectrometry and a proteome profiler. Human primary CD34+-derived endothelial cells (CD34+-ECs) were used for designing angiogenesis studies (proliferation, migration, and tubulogenesis) or in vitro models of EC monolayer (confluent monolayer ECs—CMECs) and blood–brain barrier (BBB; brain-like ECs—BLECs). Cells were treated with scCM (5 μg/mL) or protein-free endothelial basal medium (scEBM—control). CMECs or BLECs were exposed (6 h) to oxygen–glucose deprivation (OGD) conditions (1% oxygen and glucose-free medium) or normoxia (control—5% oxygen, 1 g/L of glucose) and treated with scCM or scEBM during reoxygenation (24 h). Results The analysis of different scCM batches showed a good reproducibility in terms of protein yield and composition. scCM increased CD34+-EC proliferation, tubulogenesis, and migration compared to the control (scEBM). The proteomic analysis of scCM revealed the presence of growth factors and molecules modulating cell metabolism and inflammatory pathways. Further, scCM decreased the permeability of CMECs and upregulated the expression of the junctional proteins such as occludin, VE-cadherin, and ZO-1. Such effects were possibly mediated through the activation of the interferon pathway and a moderate downregulation of Wnt signalling. Furthermore, OGD increased the permeability of both CMECs and BLECs, while scCM prevented the OGD-induced vascular leakage in both models. These effects were possibly mediated through the upregulation of junctional proteins and the regulation of MAPK/VEGFR2 activity. Conclusion Our results suggest that scCM promotes angiogenesis and the maturation of newly formed vessels while restoring the BBB function in ischemic conditions. In conclusion, our results highlight the possibility of using EPC-secretome as a therapeutic alternative to promote brain angiogenesis and protect from ischemia-induced vascular leakage.

show abstract

Section: Discussionmentioning

confidence: 99%

RETRACTED ARTICLE: Secretome of endothelial progenitor cells from stroke patients promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Endothelial colony forming cells (ECFCs) were generated as previously described. 13 In brief, cryopreserved PBMCs were seeded on 48-well plates until a colony was detected and were confluent enough to be passaged once in 24-well plates then in T-75 flasks. At this point, cells were cryopreserved (10% dimethylsulfoxide, 90% FCS) and termed P3.…”

Section: Methodsmentioning

confidence: 99%

“…The ECFC phenotype and functionality was confirmed as previously reported. 13 Stimulation of Endothelial Colony Forming Cells ECFCs were seeded onto a six-well plate (Corning Inc.) in medium (MV, PromoCell) supplemented with 5% FCS. Cells were stimulated with interferon-gamma (IFN-g, 500 U/ml, PeproTech, Rocky Hill, NJ) for 2, 4, and 8 d. Cells were then subjected to flow cytometry.…”

Section: Generation Of Endothelial Colony Forming Cellsmentioning

confidence: 99%

Expression of Class II Human Leukocyte Antigens on Human Endothelial Cells Shows High Interindividual and Intersubclass Heterogeneity

Béland¹,

Désy²,

Fekih

et al. 2023

JASN

Self Cite

View full text Add to dashboard Cite

Background HLA antigens are important targets of alloantibodies and allospecific T cells involved in graft rejection. Compared with research into understanding alloantibody development, little is known about the variability in expression of their ligands on endothelial cells. We hypothesized individual variability in the expression of HLA molecules.Methods We generated endothelial colony forming cell lines from human peripheral blood mononuclear cells (n539). Flow cytometry and immunofluorescence staining were used to analyze the cells, and we assessed the relationship between HLA-DQ expression and genotype. Two cohorts of kidney transplant recipients were analyzed to correlate HLA-DQ mismatches with the extent of intragraft microvascular injury.Results Large variability was observed in the expression of HLA class II antigens, not only between individuals but also between subclasses. In particular, HLA-DQ antigens had a low and heterogeneous expression, ranging from 0% to 85% positive cells. On a within-patient basis, this expression was consistent between endothelial cell colonies and antigen-presenting cells. HLA-DQ5 and -DQ6 were associated with higher levels of expression, whereas HLA-DQ7, -DQ8, and -DQ9 with lower. HLA-DQ5 mismatches among kidney transplant recipients were associated with significant increase in graft microvascular. ConclusionThese data challenge the current paradigm that HLA antigens, in particular HLA class II, are a single genetic and post-translational entity. Understanding and assessing the variability in the expression of HLA antigens could have clinical monitoring and treatment applications in transplantation, autoimmune diseases, and oncology.

show abstract

“…The ECFCs exhibited a cobblestone morphology and displayed the characteristic monolayer growth pattern observed in mature endothelial cells when reaching confluence. These cells were identified as adherent cells that were positive for surface markers specific to endothelial cells and hematopoietic stem cells, including acetylated low-density lipoprotein (Dil-AcLDL), eNOS, Von Willebrand factor (vWF), CD31, CD34, VE-cadherin, and kinase insert domain receptor (KDR), as described previously [17]. The fluorescent images were recorded under a laser scanning confocal microscope.…”

Section: Human Epc Isolation and Cultivationmentioning

confidence: 99%

Granulocyte Colony-Stimulating Factor Improves Endothelial Progenitor Cell-Mediated Neovascularization in Mice with Chronic Kidney Disease

Tang,

Lee,

Tseng

et al. 2023

Pharmaceutics

View full text Add to dashboard Cite

Patients with chronic kidney disease (CKD) have a higher prevalence of peripheral arterial disease (PAD), and endothelial progenitor cells (EPCs) play a pivotal role. We examined the impact of granulocyte colony-stimulating factor (G-CSF) on EPC function in response to tissue ischemia. Eight-week-old male C57BL/6J male mice were divided into sham operation and subtotal nephrectomy (SNx) groups, received hindlimb ischemic operation after seven weeks, then randomly received G-CSF or PBS intervention for four weeks with weekly follow-ups. SNx mice had significantly reduced limb reperfusion, decreased plasma EPC mobilization, and impaired angiogenesis in ischemic hindlimbs compared to the control group. However, G-CSF increased IL-10 and reversed these adverse changes. Additionally, ischemia-associated protein expressions, including IL-10, phospho-STAT3, VEGF, and phospho-eNOS, were significantly downregulated in the ischemic hindlimbs of SNx mice versus control, but these trends were reversed by G-CSF. Furthermore, in cultured EPCs, G-CSF significantly attenuated the decrease in EPC function initiated by indoxyl sulfate through IL-10. Overall, we discovered that G-CSF can improve EPC angiogenic function through a hypoxia/IL-10 signaling cascade and impede neovascular growth in response to ischemia of SNx mice. Our results highlight G-CSF’s potential to restore angiogenesis in CKD patients with PAD via EPC-based methods.

show abstract

Endothelial colony forming cells generated from cryopreserved peripheral blood mononuclear cells

Cited by 3 publications

References 29 publications

RETRACTED ARTICLE: Secretome of endothelial progenitor cells from stroke patients promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage

RETRACTED ARTICLE: Secretome of endothelial progenitor cells from stroke patients promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage

Expression of Class II Human Leukocyte Antigens on Human Endothelial Cells Shows High Interindividual and Intersubclass Heterogeneity

Granulocyte Colony-Stimulating Factor Improves Endothelial Progenitor Cell-Mediated Neovascularization in Mice with Chronic Kidney Disease

Contact Info

Product

Resources

About