Comparison between total endothelial progenitor cell isolation versus enriched Cd133+ culture

Amelia Casamassimi, Maria Luisa Balestrieri, Carmela Fiorito, Concetta Schiano, Ciro Maione, Raffaele Rossiello, Vincenzo Grimaldi, Vincenzo Del Giudice, Ciro Balestrieri, Bartolomeo Farzati, Vincenzo Sica, Claudio Napoli

Research output: Contribution to journalArticle

Abstract

Endothelial progenitor cells (EPCs) play a role in endogenous neovascularization of ischaemic tissues. Isolation and characterization of EPCs from circulating mononuclear cells are important for developing targeted cellular therapies and reproducibility of data are the major scientific goals. Here we compared two currently employed isolation methods, i.e. from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133+ cells, by defining the cell morphology and functional activities. We show that EPCs from cultured PBMCs resulted in an adherent population of 23% ± 4% merged cells positive for Dil-Ac-LDL and lectin, whereas the percentage of double positive cells in cultured CD133+ enriched cells was 50% ± 7% (P <0.01). These data were obtained through a novel and a more complete method of analysis of cell calculations (specifically by dividing each microscope field into 120 subfields). When stimulated with tumour necrosis factor α (TNF)-α and glucose, cell number was reduced in EPCs from total PBMCs and, more consistently, in CD133+ enriched cells. However, both cultured total PBMCs and CD133+ enriched cells respond similarly to TNF-α or glucose-induced p38-phosphorylation.EPCs from both procedures show similar results in terms of phenotype and response to modulators of their functional activities. However, when the cell phenotype of CD133 + enrichment-derived cells was compared with that of cells from the total PBMC, a significant increase in CD133+ expression was observed (P <0.01) This may have relevance during intervention studies using cultured EPCs.

Original languageEnglish
Pages (from-to)503-511
Number of pages9
JournalJournal of Biochemistry
Volume141
Issue number4
DOIs
Publication statusPublished - Apr 2007

Fingerprint

Cell Separation
Endothelial cells
Cell culture
Blood
Blood Cells
Tumor Necrosis Factor-alpha
Glucose
Phosphorylation
Lectins
Cultured Cells
Modulators
Endothelial Progenitor Cells
Microscopes
Cells
Phenotype
Tissue
Cell Count

Keywords

  • CD133
  • Endothelial progenitor cells
  • Glucose
  • TNF

ASJC Scopus subject areas

  • Biochemistry

Cite this

Comparison between total endothelial progenitor cell isolation versus enriched Cd133+ culture. / Casamassimi, Amelia; Balestrieri, Maria Luisa; Fiorito, Carmela; Schiano, Concetta; Maione, Ciro; Rossiello, Raffaele; Grimaldi, Vincenzo; Del Giudice, Vincenzo; Balestrieri, Ciro; Farzati, Bartolomeo; Sica, Vincenzo; Napoli, Claudio.

In: Journal of Biochemistry, Vol. 141, No. 4, 04.2007, p. 503-511.

Research output: Contribution to journalArticle

Casamassimi, A, Balestrieri, ML, Fiorito, C, Schiano, C, Maione, C, Rossiello, R, Grimaldi, V, Del Giudice, V, Balestrieri, C, Farzati, B, Sica, V & Napoli, C 2007, 'Comparison between total endothelial progenitor cell isolation versus enriched Cd133+ culture', Journal of Biochemistry, vol. 141, no. 4, pp. 503-511. https://doi.org/10.1093/jb/mvm060
Casamassimi, Amelia ; Balestrieri, Maria Luisa ; Fiorito, Carmela ; Schiano, Concetta ; Maione, Ciro ; Rossiello, Raffaele ; Grimaldi, Vincenzo ; Del Giudice, Vincenzo ; Balestrieri, Ciro ; Farzati, Bartolomeo ; Sica, Vincenzo ; Napoli, Claudio. / Comparison between total endothelial progenitor cell isolation versus enriched Cd133+ culture. In: Journal of Biochemistry. 2007 ; Vol. 141, No. 4. pp. 503-511.
@article{2514419fff0445febbb1cee9488d06f0,
title = "Comparison between total endothelial progenitor cell isolation versus enriched Cd133+ culture",
abstract = "Endothelial progenitor cells (EPCs) play a role in endogenous neovascularization of ischaemic tissues. Isolation and characterization of EPCs from circulating mononuclear cells are important for developing targeted cellular therapies and reproducibility of data are the major scientific goals. Here we compared two currently employed isolation methods, i.e. from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133+ cells, by defining the cell morphology and functional activities. We show that EPCs from cultured PBMCs resulted in an adherent population of 23{\%} ± 4{\%} merged cells positive for Dil-Ac-LDL and lectin, whereas the percentage of double positive cells in cultured CD133+ enriched cells was 50{\%} ± 7{\%} (P <0.01). These data were obtained through a novel and a more complete method of analysis of cell calculations (specifically by dividing each microscope field into 120 subfields). When stimulated with tumour necrosis factor α (TNF)-α and glucose, cell number was reduced in EPCs from total PBMCs and, more consistently, in CD133+ enriched cells. However, both cultured total PBMCs and CD133+ enriched cells respond similarly to TNF-α or glucose-induced p38-phosphorylation.EPCs from both procedures show similar results in terms of phenotype and response to modulators of their functional activities. However, when the cell phenotype of CD133 + enrichment-derived cells was compared with that of cells from the total PBMC, a significant increase in CD133+ expression was observed (P <0.01) This may have relevance during intervention studies using cultured EPCs.",
keywords = "CD133, Endothelial progenitor cells, Glucose, TNF",
author = "Amelia Casamassimi and Balestrieri, {Maria Luisa} and Carmela Fiorito and Concetta Schiano and Ciro Maione and Raffaele Rossiello and Vincenzo Grimaldi and {Del Giudice}, Vincenzo and Ciro Balestrieri and Bartolomeo Farzati and Vincenzo Sica and Claudio Napoli",
year = "2007",
month = "4",
doi = "10.1093/jb/mvm060",
language = "English",
volume = "141",
pages = "503--511",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - Comparison between total endothelial progenitor cell isolation versus enriched Cd133+ culture

AU - Casamassimi, Amelia

AU - Balestrieri, Maria Luisa

AU - Fiorito, Carmela

AU - Schiano, Concetta

AU - Maione, Ciro

AU - Rossiello, Raffaele

AU - Grimaldi, Vincenzo

AU - Del Giudice, Vincenzo

AU - Balestrieri, Ciro

AU - Farzati, Bartolomeo

AU - Sica, Vincenzo

AU - Napoli, Claudio

PY - 2007/4

Y1 - 2007/4

N2 - Endothelial progenitor cells (EPCs) play a role in endogenous neovascularization of ischaemic tissues. Isolation and characterization of EPCs from circulating mononuclear cells are important for developing targeted cellular therapies and reproducibility of data are the major scientific goals. Here we compared two currently employed isolation methods, i.e. from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133+ cells, by defining the cell morphology and functional activities. We show that EPCs from cultured PBMCs resulted in an adherent population of 23% ± 4% merged cells positive for Dil-Ac-LDL and lectin, whereas the percentage of double positive cells in cultured CD133+ enriched cells was 50% ± 7% (P <0.01). These data were obtained through a novel and a more complete method of analysis of cell calculations (specifically by dividing each microscope field into 120 subfields). When stimulated with tumour necrosis factor α (TNF)-α and glucose, cell number was reduced in EPCs from total PBMCs and, more consistently, in CD133+ enriched cells. However, both cultured total PBMCs and CD133+ enriched cells respond similarly to TNF-α or glucose-induced p38-phosphorylation.EPCs from both procedures show similar results in terms of phenotype and response to modulators of their functional activities. However, when the cell phenotype of CD133 + enrichment-derived cells was compared with that of cells from the total PBMC, a significant increase in CD133+ expression was observed (P <0.01) This may have relevance during intervention studies using cultured EPCs.

AB - Endothelial progenitor cells (EPCs) play a role in endogenous neovascularization of ischaemic tissues. Isolation and characterization of EPCs from circulating mononuclear cells are important for developing targeted cellular therapies and reproducibility of data are the major scientific goals. Here we compared two currently employed isolation methods, i.e. from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133+ cells, by defining the cell morphology and functional activities. We show that EPCs from cultured PBMCs resulted in an adherent population of 23% ± 4% merged cells positive for Dil-Ac-LDL and lectin, whereas the percentage of double positive cells in cultured CD133+ enriched cells was 50% ± 7% (P <0.01). These data were obtained through a novel and a more complete method of analysis of cell calculations (specifically by dividing each microscope field into 120 subfields). When stimulated with tumour necrosis factor α (TNF)-α and glucose, cell number was reduced in EPCs from total PBMCs and, more consistently, in CD133+ enriched cells. However, both cultured total PBMCs and CD133+ enriched cells respond similarly to TNF-α or glucose-induced p38-phosphorylation.EPCs from both procedures show similar results in terms of phenotype and response to modulators of their functional activities. However, when the cell phenotype of CD133 + enrichment-derived cells was compared with that of cells from the total PBMC, a significant increase in CD133+ expression was observed (P <0.01) This may have relevance during intervention studies using cultured EPCs.

KW - CD133

KW - Endothelial progenitor cells

KW - Glucose

KW - TNF

UR - http://www.scopus.com/inward/record.url?scp=34347396894&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34347396894&partnerID=8YFLogxK

U2 - 10.1093/jb/mvm060

DO - 10.1093/jb/mvm060

M3 - Article

C2 - 17308344

AN - SCOPUS:34347396894

VL - 141

SP - 503

EP - 511

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 4

ER -