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The Promoting Effect of Vascular Endothelial Growth Factor on the Generation of CD34+ Cells from Mouse Embryonic Stem Cells
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Abstract:
Objective:To study the promoting effect of vascular endothelial growth factor (VEGF) on the generation of CD34+ cells from mouse embryonic stem cell(ESC) in vitro.Methods: ES-D3 cells were allowed to grow on mouse fetal fibroblast feeder layer to form embryoid bodies(EB), then EB cells were transferred into medium supplemented with different concentration of VEGF and VEGF+SCF for 1 week. Six groups were designed , i.e. VEGF 5 μg/L, VEGF 10 μg/L, VEGF 20 μg/L, VEGF 5 μg/L＋SCF, VEGF 10 μg/L＋SCF and VEGF 20 μg/L＋SCF. The group of spontaneous differentiation without cytokine(s) was served as control. The expression of CD34 mRNA was detected by RT-PCR. The content of CD34+ cells in each group was measured by flow cytometry. The cells derived from ESC were incubated in semisolid methycellulose cultures. The numbers of total colony-forming units in culture(CFU-C) were counted by reverse microscope. Results：ES-D3 grew and formed EBs at day 4. VEGF gave the stimulatory effects on the expression of CD34 mRNA, the generation of CD34+ cells and CFU-C in EB as a single factor . The effects of VEGF＋SCF were the most effective in the experimental groups according to the percent of CD34+ cells and the numbers of hematopoietic colonies. The most highest inducing efficacy were achieved when VEGF 20 μg/L or VEGF 10 μg/L combined with SCF were used.Conclusion: VEGF can promote the differentiation of ESC into CD34+ cells in vitro. The strongest effects were achieved when VEGF was used in combination with SCF.
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