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 language | English |
|---|---|
| Pages (from-to) | 503-511 |
| Number of pages | 9 |
| Journal | Journal of Biochemistry |
| Volume | 141 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Apr 2007 |
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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 journal › Article
}
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 -