Human PBMNCs were isolated as described previously, and 1 × 103 PBMNCs were added to each well of a 24-well plate (BD Falcon, Bedford, MA, USA) and cultured in serum-free QQc culture medium for 7 d17 (link)–19 (link, link) . Briefly, QQc culture medium is an optimized combination of growth factors and cytokines (20 ng/mL thrombopoietin, 20 ng/mL interleukin (IL) 6, 100 ng/mL stem cell factor (SCF), 100 ng/mL Flt-3 ligand, and 50 ng/mL vascular endothelial growth factor; all from Peprotech, Rocky Hills, NJ, USA) in serum-free stem cell medium (Stemcell Technologies, Vancouver, Canada). QQc culture was performed in this medium for 7 d, which has been shown to dramatically and optimally expand and enhance the vasculogenic potential of EPCs.
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Enhancing Vasculogenic Potential of EPCs
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Cytokines
Epcs
Flt 3 ligand
Growth factors
Human
Interleukin 6
Serum
Stem cell factor
Stemcell
Thrombopoietin
Vascular endothelial growth factor
Corresponding Organization : Shonan Kamakura General Hospital
Other organizations : The University of Tokyo, Juntendo University
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Variable analysis
independent variables
- Isolation of PBMNCs
- Addition of 1 × 10^3 PBMNCs to each well of a 24-well plate
- Culture of PBMNCs in serum-free QQc culture medium for 7 days
- Expansion and enhancement of the vasculogenic potential of EPCs
- Serum-free QQc culture medium composition (20 ng/mL thrombopoietin, 20 ng/mL interleukin (IL) 6, 100 ng/mL stem cell factor (SCF), 100 ng/mL Flt-3 ligand, and 50 ng/mL vascular endothelial growth factor)
- Culture duration of 7 days
- No positive or negative controls were explicitly mentioned in the provided information.
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