Abstract 4297: Effect of Human Collagen I and Matrigel Matrices on Human Embryonic Stem Cell Derived Endothelial Cells
Introduction: Human embryonic stem cell-derived endothelial cells (hESC-ECs) show potential promise as regenerative therapies for cardiovascular diseases. We hypothesize extracellular matrix and soluble factors-human Collagen I + Matrigel (C/M) matrices can improve survival, proliferation, and angiogenesis of hESC-ECs.
Methods: A novel double fusion (DF) reporter gene that consists of β-actin promoter driving firefly luciferase and enhanced green fluorescence protein was stably transduced into hESCs (H9) using a self-inactivating lentiviral vector. Floating embryoid body formation was performed to induce ECs differentiation. HESC-ECs + C/M mixture was seeded on a Flex Cell tissue culture system. To confirm effect, morphology and phenotype, bioluminescence (BLI) imaging, H&E staining and immunostaining were performed on these constructs.
Results: After 12 days of differentiation, ~2.4±0.3% CD31+ cells were isolated and expanded in vitro. These cells express ~96.2±2% CD31, ~91.3±4% VE-cadherin, and behave similarly to HUVECs (Figure 1⇓). BLI imaging results indicate C/M matrices can significantly improve hESC-ECs proliferation (220±25%) compared with medium alone (140±12%) or Collagen I matrix alone (180±18%) (P<0.05 for both). After growing within the C/M construct for 15 days, hESC-ECs maintained expression of their endothelial markers (~95.6±1% CD31 and ~90.2±3% VE-cadherin).
Conclusion: This is the first study to demonstrate the beneficial effects of C/M matrices on hESC-ECs proliferation in vitro. Confirmation of their effect on transplanted hESC-ECs in vivo using molecular imaging is currently in process.