Standardization of methods to quantify and culture endothelial colony‐forming cells derived from peripheral blood

Smadja, David M.; Melero‐Martin, Juan M.; Eikenboom, Jeroen; Bowman, Mackenzie; Sabatier, Florence; Randi, Anna M.

doi:10.1111/jth.14462

Cited by 58 publications

(41 citation statements)

References 36 publications

(44 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, the authors believe that age has not had an in uence, as previously reported in previous studies. [11] In this study, the ECFC differentiation and proliferation capacity was not impaired in GPA patients, in contrast to the ndings of Wilde et al The authors point out that a patient with active disease had a much higher number of colony, which raised the average of the GPA group. In the present study, a speci c group of patients, all with PR3-ANCA, was evaluated.…”

Section: Discussioncontrasting

confidence: 79%

“…[9] EPC isolated from the peripheral blood generate endothelial colony-forming cells (ECFC) in vitro, which have a stable culture-wide phenotype and high proliferative capacity; therefore, these cells are the most commonly used in in vitro studies. [10,11] Although it is known that endothelial damage has a key effect on AAV, little has been studied about vascular repair. Previous studies suggest that endothelial repair dysfunction in AAV patients may be due to changes in circulating EPC.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Angiogenic properties of human endothelial colony-forming cells in Granulomatosis with Polyangiitis

Rio¹,

Prieto²,

Frade³

et al. 2020

Preprint

View full text Add to dashboard Cite

BACKGROUND Endothelial progenitor cells are essential for vascular homeostasis. Considering the recurrent nature of granulomatosis with polyangiitis (GPA) the aim of the study was to evaluate the angiogenic capacity of endothelial colony-forming cells (ECFC), which have the capacity for neovasculogenesis in vitro, of the patients with GPA, before and after plasma stimulation. METHODS Thirteen GPA PR3-positive patients and 15 healthy controls were included. ECFC were isolated from periferic blood and characterized by flow cytometry (FACS). Capillary tube formation (Matrigel assay) and scratching assay were measured during 24 hours. The migration assay was also performed after overnight incubation with healthy control plasma and active GPA patient plasma. Three patients with active disease where submitted to recollection of ECFC after treatment for longitudinal evaluation. RESULTS The culture was successful in 62% of GPA patients and 57% of controls. In the matrigel assay the ECFC from patients and controls showed similar capillary structures formation, however ECFC from inactive GPA alone showed early loss of angiogenic capacity between 15 and 24 hours. In scratching assay, there was an impairment in the proliferative capacity of the ECFC between GPA patients and controls without significant difference (12th hour, p = 0.05). When incubated with control plasma, ECFC of remission GPA patients showed a significant lower migration capacity after the 4th hour (p = 0.0001). In longitudinal analysis, ECFC isolated after treatment showed significantly lower migration rates. CONCLUSIONS PR3-positive remission GPA ECFC demonstrated early loss of tube formation and less proliferative capacity compared to those of healthy controls, suggesting impairment of endothelial function.

show abstract

Section: Discussioncontrasting

confidence: 79%

Section: Introductionmentioning

confidence: 99%

Angiogenic properties of human endothelial colony-forming cells in Granulomatosis with Polyangiitis

Rio¹,

Prieto²,

Frade³

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…However, this cell population can be challenged since no correlation has been reported between the number of these circulating cells and the number of endothelial colonies forming cells (ECFCs) obtained in cell culture [16,17]. Of note, ECFCs are considered by [18]. In a recent meta-analysis reporting pooled data on the association between levels of circulating progenitor cells, cardiovascular outcomes and death, CD34 + and CD34 + CD133 + were the most frequent cell population associated with cardiovascular outcomes and death.…”

Section: Discussionmentioning

confidence: 99%

Multidimensional Proteomic Approach of Endothelial Progenitors Demonstrate Expression of KDR Restricted to CD19 Cells

Guerin

Guyonnet

Goudot

et al. 2020

Stem Cell Rev and Rep

Self Cite

View full text Add to dashboard Cite

Endothelial progenitor cells (EPCs) are involved in vasculogenesis and cardiovascular diseases. However, the phenotype of circulating EPCs remains elusive but they are more often described as CD34 + KDR + . The aim of the study was to extensively characterize circulating potential vasculogenic stem cell candidates in two populations of patients with cardiovascular disease by powerful multidimensional single cell complementary cytometric approaches (mass, imaging and flow). We identified cellular candidates in one patient before and after bioprosthetic total artificial heart implantation and results were confirmed in healthy peripheral and cord blood by mass cytometry. We also quantified cellular candidates in 10 patients with different COVID-19 severity. Both C-TAH implantation and COVID-19 at critical stage induce a redistribution of circulating CD34 + and CD19 + sub-populations in peripheral blood. After C-TAH implantation, circulating CD34 + progenitor cells expressed c-Kit stem marker while specific subsets CD34 + CD133 −/+ CD45 −/dim c-Kit + KDR − were mobilized. KDR was only expressed by CD19 + B-lymphocytes and CD14 + monocytes subpopulations in circulation. We confirmed by mass cytometry this KDR expression on CD19 + in healthy peripheral and cord blood, also with a VE-cadherin expression, confirming absence of endothelial lineage marker on CD34 + subtypes. In COVID-19, a significant mobilization of CD34 + c-Kit + KDR − cells was observed between moderate and critical COVID-19 patients regardless CD133 or CD45 expression. In order to better evaluate EPC phenotype, we performed imaging flow cytometry measurements of immature CD34 + KDR + cells in cord blood and showed that, after elimination of non-circular events, those cells were all CD19 + . During COVID-19, a significant mobilization of CD19 + KDR + per million of CD45 + cells was observed between moderate and critical COVID-19 patients regardless of CD34 expression. CD34 + c-Kit + cells are mobilized in both cardiovascular disease described here. KDR cells in peripheral blood are CD19 positive cells and are not classic vasculogenic stem and/or progenitor cells. A better evaluation of c-Kit and KDR expressing cells will lead to the redefinition of circulating endothelial progenitors. Graphical abstract Central illustration figure. Multidimensional proteomic approach of endothelial progenitors demonstrate expression of KDR restricted to CD19 cells. ...

show abstract

“…MACs originate from the myeloid–monocytic lineage and support endothelial repair and vascular regeneration through largely paracrine signals [ 18 ]. In turn, ECFCs represent the only known truly endothelial precursor, as they lack hematopoietic markers, display high clonogenic potential, are able to aggregate into bidimensional tube networks in vitro and to originate patent vessels in vivo [ 19 , 20 ]. Therefore, ECFCs provide the most promising cellular candidate for therapeutic purposes by physically engrafting within injured endothelial and also delivering pro-angiogenic cues to adjacent endothelial cells [ 21 ].…”

Section: Introductionmentioning

confidence: 99%

Therapeutic Potential of Endothelial Colony-Forming Cells in Ischemic Disease: Strategies to Improve their Regenerative Efficacy

Faris

Negri

Perna

et al. 2020

IJMS

View full text Add to dashboard Cite

Cardiovascular disease (CVD) comprises a range of major clinical cardiac and circulatory diseases, which produce immense health and economic burdens worldwide. Currently, vascular regenerative surgery represents the most employed therapeutic option to treat ischemic disorders, even though not all the patients are amenable to surgical revascularization. Therefore, more efficient therapeutic approaches are urgently required to promote neovascularization. Therapeutic angiogenesis represents an emerging strategy that aims at reconstructing the damaged vascular network by stimulating local angiogenesis and/or promoting de novo blood vessel formation according to a process known as vasculogenesis. In turn, circulating endothelial colony-forming cells (ECFCs) represent truly endothelial precursors, which display high clonogenic potential and have the documented ability to originate de novo blood vessels in vivo. Therefore, ECFCs are regarded as the most promising cellular candidate to promote therapeutic angiogenesis in patients suffering from CVD. The current briefly summarizes the available information about the origin and characterization of ECFCs and then widely illustrates the preclinical studies that assessed their regenerative efficacy in a variety of ischemic disorders, including acute myocardial infarction, peripheral artery disease, ischemic brain disease, and retinopathy. Then, we describe the most common pharmacological, genetic, and epigenetic strategies employed to enhance the vasoreparative potential of autologous ECFCs by manipulating crucial pro-angiogenic signaling pathways, e.g., extracellular-signal regulated kinase/Akt, phosphoinositide 3-kinase, and Ca2+ signaling. We conclude by discussing the possibility of targeting circulating ECFCs to rescue their dysfunctional phenotype and promote neovascularization in the presence of CVD.

show abstract

Standardization of methods to quantify and culture endothelial colony‐forming cells derived from peripheral blood

Cited by 58 publications

References 36 publications

Angiogenic properties of human endothelial colony-forming cells in Granulomatosis with Polyangiitis

Angiogenic properties of human endothelial colony-forming cells in Granulomatosis with Polyangiitis

Multidimensional Proteomic Approach of Endothelial Progenitors Demonstrate Expression of KDR Restricted to CD19 Cells

Therapeutic Potential of Endothelial Colony-Forming Cells in Ischemic Disease: Strategies to Improve their Regenerative Efficacy

Contact Info

Product

Resources

About