Isolating and expanding endothelial progenitor cells from peripheral blood on peptide‐functionalized polystyrene surfaces

Elkhodiry, Mohamed A; Boulanger, Mariève D.; Bashth, Omar; Tanguay, Jean‐François; Laroche, Gaétan; Hoesli, Corinne A.

doi:10.1002/bit.27107

Cited by 5 publications

(9 citation statements)

References 39 publications

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“…Out of the four conditions, only the immobilized anti-CD144 on the conjugated protein G had a significant effect in enhancing ECFC capture under flow (Figure 6). Surfaces with adsorbed anti-CD144 showed no significant difference in the number of captured ECFCs compared with surfaces modified with anti-CD14, a surface antigen that is not expressed by ECFCs 27 . Surfaces coated with rat tail collagen, a commonly used ECFC substrate, also had a significantly lower number of captured cells compared to the immobilized anti-CD144 on the conjugated protein G. As a control, PBMCs, rich in CD14+ cells (>45%) but with low or undetectable CD144+ cell populations (<0.1%), were separately circulated in the same conditions over the same surfaces.…”

Section: Antibody-functionalized Surfaces Can Capture Ecfcsmentioning

confidence: 85%

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Bioaffinity-based surface-immobilization of antibodies to capture endothelial colony-forming cells

Boulanger

Elkhodiry

Bashth

et al. 2021

Preprint

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Maximizing the re-endothelialization of vascular implants such as prostheses or stents has the potential to significantly improve their long-term performance. Endothelial progenitor cell capture stents with surface-immobilized antibodies show significantly improved endothelialization in the clinic. However, most current antibody-based stent surface modification strategies rely on antibody adsorption or direct conjugation via amino or carboxyl groups which leads to poor control over antibody surface concentration and/or molecular orientation, and ultimately bioavailability for cell capture. Here, we assess the utility of a bioaffinity-based surface modification strategy consisting of a surface-conjugated cysteine-tagged protein G molecules that immobilize Immunoglobulin G (IgG) antibodies via the Fc domain to capture circulating endothelial colony-forming cells (ECFCs). The cysteine-tagged protein G was grafted onto aminated substrates at different concentrations as detected by an enzyme-linked immunosorbent assay and fluorescence imaging. Different IgG antibodies were successfully immobilized on the protein G-modified surfaces and higher antibody surface concentrations were achieved compared to passive adsorption methods. Surfaces with immobilized antibodies targeting endothelial surface proteins, such as CD144, significantly enhanced the capture of circulating ECFCs in vitro compared to surfaces with non-endothelial specific antibodies such as anti-CD14. This work presents a potential avenue for enhancing the clinical performance of vascular implants by using covalent grafting of protein G to immobilize IgG antibodies more effectively.

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Section: Antibody-functionalized Surfaces Can Capture Ecfcsmentioning

confidence: 85%

“…Peripheral blood mononuclear cells (PBMCs) were isolated from fresh adult human peripheral blood and ECFCs were expanded as previously described 27 . Fresh blood samples were collected from adult donors under informed consent following a protocol (Study No.…”

Section: Ecfc Capture Under Flowmentioning

confidence: 99%

Bioaffinity-based surface-immobilization of antibodies to capture endothelial colony-forming cells

Boulanger

Elkhodiry

Bashth

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Out of the four conditions, only the immobilized anti-CD144 on the conjugated protein G polypeptide had a significant effect in enhancing ECFC capture under flow ( Fig 5 ). Surfaces with adsorbed anti-CD144 showed no significant difference in the number of captured ECFCs compared with surfaces modified with anti-CD14, a surface antigen that is not expressed by ECFCs [ 30 ]. Surfaces coated with rat tail collagen, a commonly used ECFC substrate, also had a significantly lower number of captured cells compared to the immobilized anti-CD144 on the conjugated protein G polypeptide.…”

Section: Resultsmentioning

confidence: 99%

“…Peripheral blood mononuclear cells (PBMCs) were isolated from fresh adult human peripheral blood and ECFCs were expanded as previously described [ 30 ]. Fresh blood samples were collected from adult donors (N = 4: 2 females and 2 males; mean age 25.5 years) under informed consent following a protocol (Study No.…”

Section: Methodsmentioning

confidence: 99%

Bioaffinity-based surface immobilization of antibodies to capture endothelial colony-forming cells

et al. 2022

Self Cite

View full text Add to dashboard Cite

Maximizing the re-endothelialization of vascular implants such as prostheses or stents has the potential to significantly improve their long-term performance. Endothelial progenitor cell capture stents with surface-immobilized antibodies show significantly improved endothelialization in the clinic. However, most current antibody-based stent surface modification strategies rely on antibody adsorption or direct conjugation via amino or carboxyl groups which leads to poor control over antibody surface concentration and/or molecular orientation, and ultimately bioavailability for cell capture. Here, we assess the utility of a bioaffinity-based surface modification strategy to immobilize antibodies targeting endothelial cell surface antigens. A cysteine-tagged truncated protein G polypeptide containing three Fc-binding domains was conjugated onto aminated polystyrene substrates via a bi-functional linking arm, followed by antibody immobilization. Different IgG antibodies were successfully immobilized on the protein G-modified surfaces. Covalent grafting of the protein G polypeptide was more effective than surface adsorption in immobilizing antibodies at high density based on fluorophore-labeled secondary antibody detection, as well as endothelial colony-forming cell capture through anti-CD144 antibodies. This work presents a potential avenue for enhancing the performance of cell capture strategies by using covalent grafting of protein G polypeptides to immobilize IgG antibodies.

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“…Human peripheral blood (PB) contains a variety of stem cells, including mononuclear cells (MNCs) and hematopoietic stem/progenitor cells (HSPCs) from the bone marrow, [ 1 ] mesenchymal stem cells (MSCs), [ 2 , 3 ] endothelial progenitor cells (EPCs), [ 4 ] and very small embryonic-like stem cells (VSELs). [ 5 , 6 ] These cells express capacities of differentiation and regeneration with HSPCs/MNCs differentiating into hepatocyte, [ 7 ] and MSCs exhibiting osteogenesis and adipose differentiation.…”

Section: Introductionmentioning

confidence: 99%

Culture of leukocyte-derived cells from human peripheral blood: Increased expression of pluripotent genes OCT4, NANOG, SOX2, self-renewal gene TERT and plasticity

et al. 2023

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There are few stem cells in human peripheral blood (PB). Increasing the population and plasticity of stem cells in PB and applying it to regenerative medicine require suitable culture methods. In this study, leukocyte populations 250 mL of PB were collected using a blood separator before that were cultured in optimal cell culture medium for 4 to 7 days. After culturing, stemness characteristics were analyzed, and red blood cells were removed from the cultured cells. In our results, stemness markers of the leukocyte populations Sca-1 + CD45 + , CD117 + CD45 + , and very small embryonic-like stem cells CD34 + Lin − CD45 − and CXCR4 + Lin − CD45 − were significantly increased. Furthermore, the expression of stem cell genes OCT4 (POU5F1), NANOG, SOX2, and the self-renewal gene TERT was analyzed by quantitative real-time polymerase chain reaction in these cells, and it showed a significant increase. These cells could be candidates for multi-potential cells and were further induced using transdifferentiation culture methods. These cells showed multiple differentiation potentials for osteocytes, nerve cells, cardiomyocytes, and hepatocytes. These results indicate that appropriate culture methods can be applied to increase expression of pluripotent genes and plasticity. Leukocytes of human PB can be induced to trans-differentiate into pluripotent potential cells, which will be an important breakthrough in regenerative medicine.

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Isolating and expanding endothelial progenitor cells from peripheral blood on peptide‐functionalized polystyrene surfaces

Cited by 5 publications

References 39 publications

Bioaffinity-based surface-immobilization of antibodies to capture endothelial colony-forming cells

Bioaffinity-based surface-immobilization of antibodies to capture endothelial colony-forming cells

Bioaffinity-based surface immobilization of antibodies to capture endothelial colony-forming cells

Culture of leukocyte-derived cells from human peripheral blood: Increased expression of pluripotent genes OCT4, NANOG, SOX2, self-renewal gene TERT and plasticity

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