Abstract 17999: Synthetic HGF-Mimetic Induces Endothelial Phenotype in Wharton’s Jelly Mesenchymal Stem Cells and Promotes Angiogenesis
Introduction: Although hepatocyte growth factor (HGF) has been shown to increase capillary density, short half-life remains a major limiting factor for therapeutic use. The potential utility of umbilical cord Wharton’s jelly mesenchymal stem cells (WJ-MSCs) for vascular regeneration remains unknown.
Hypothesis: A synthetic boron-based (phthalazinone and oxazine derivatives) HGF-mimetic (HGF-m) small molecule would promote endothelial differentiation of WJ-MSCs.
Methods: WJ-MSCs were cultured in absence/presence of HGF-m (0.01, 0.05 and 0.10 μmol). Flow cytometry was performed at the 48 h and 96 h for CD31 and CD34 expression. To confirm our findings and to elucidate the underlying mechanism involved, we performed Western blot analysis using protein obtained from WJ-MSCs. In vivo Matrigel plug assay, using control or HGF-m pretreated WJ-MSCs was used to test angiogenesis potential of HGF-m in vivo.
Results: Minimal expression of CD31 and CD34 was noted in control WJ-MSCs (Figure). HGF- exposure resulted in marked and dose-dependent increase in CD34+ WJ-MSCs at 96 h (61%, 83%, 83%; P<0.05 vs. controls for all comparisons) and also CD31+/CD34+ cells (11.5%, 12.4%, 16%; P<0.05 vs. controls for all comparisons) (Figure). Western analysis confirmed increased expression of CD31, CD34, p-Met and VEGF in HGF-m-treated cells. Furthermore, Western analysis showed increased expression of p-Akt, p-ERK1/2 and FLK1 in HGF-m-treated WJ-MSCs, indicating endothelial commitment and angiogenic potential. In vivo, Matrigel plug assay revealed significant and dose-dependent increase in capillary formation by HGF-m-treated WJ-MSCs (Figure).
Conclusions: A synthetic HGF-memetic small molecule can effectively enhance endothelial differentiation and angiogenic potential of WJ-MSCs both in vitro and in vivo. These findings have important translational implications for enhancing myocardial angiogenesis with cell-based therapies.
Author Disclosures: Z. Shah: None. T. Buehler: None. L. Chen: None. R. Vincent: None. B. Blount: None. A. Burnett: None. J. Hauptman: None. M. Girgis: None. B. Das: None. B. Dawn: None.
- © 2016 by American Heart Association, Inc.