An affordable method to obtain cultured endothelial cells from peripheral blood.
Bottom Line: The use of heparin, a minimum blood volume of 30 ml, fibronectin as a coating matrix and endothelial growing media-2 supplemented with 20% FBS increased the success of obtaining EPC cultures up to 80% of the processed samples while reducing EPC colony appearance mean time to a minimum of 13 days.Blood samples exhibiting higher cEPC numbers resulted in reduced EPC colony appearance mean time.Functionally, cultured EPC showed decreased growing and vasculogenic capacity when compared to HUVEC.
Affiliation: Research Foundation, Hospital Clínico of Valencia - INCLIVA, Valencia, Spain.Show MeSH
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Mentions: Endothelial progenitor cells and HUVEC growth curves are presented in Figure 5A. Cells were counted every day until cells reached confluence. Endothelial progenitor cells showed longer lag phase (2.4 days) compared with HUVEC (1.3 days; P < 0.05). Endothelial progenitor cells cultures were found to have shorter exponential growth phase. Endothelial progenitor cells plateau growth phase was reached on the 4th day after seeding at a saturation density of 5.2 × 105 cells/cm2. By contrast, HUVEC reached plateau growth phase on the 5th day after seeding at a saturation density of 17.7 × 105 cells/cm2 (P < 0.001; Fig. 5A).
Affiliation: Research Foundation, Hospital Clínico of Valencia - INCLIVA, Valencia, Spain.