Abstract 20680: Human Endothelial Cells Derived from Induced Pluripotent Stem Cells Enhance Perfusion in a Murine Model of Peripheral Arterial Disease
Stem cell therapy for angiogenesis and vascular regeneration has been investigated using diverse cell sources, including adult stem cells and embryonic stem cells. In the present study, we investigated the potential of endothelial cells (ECs) derived from human induced pluripotent stem cells (iPSCs) in promoting the perfusion of ischemic tissue in a murine model of peripheral arterial disease. The iPSCs were generated from human adult dermal fibroblasts using retrovirus-mediated transduction of Oct-4, Sox-2, Klf-4 and c-Myc. The iPSCs expressed pluripotency markers such as SSEA3/4 and alkaline phosphatase and formed teratomas in vivo. Endothelial lineage differentiation was initiated by culturing iPSCs for 14 days in differentiation media supplemented with BMP-4 and VEGF. CD144-positive iPSC-ECs were isolated using fluorescence activated cell sorting and were characterized by gene expression analysis, immunofluorescence staining and functional assays to confirm their endothelial identity. These purified iPSC-ECs exhibited endothelial angiogenic behavior by forming capillary-like structures in matrigel and incorporating acetylated-LDL. They stained positive for EC markers such as KDR, CD31, CD144 and eNOS. iPSC-ECs were stably transduced with a double fusion construct encoded by the ubiquitin promoter, firefly luciferase for bioluminescence imaging (BLI) and green fluorescence protein for fluorescent detection (pUb-Fluc-GFP). 5X105 iPSC-ECs were intramuscularly injected into SCID mice with hindlimb ischemia at day 0 and day 7 post-surgery (n=11). Bioluminescent imaging (BLI) and laser Doppler blood perfusion were performed to assess cell survival and functional improvement respectively. BLI data showed that iPSC-ECs survived in the ischemic limb throughout the 2 weeks of assessment. In addition, laser Doppler imaging showed that the ratio of blood perfusion (ischemic to normal limb) was modestly increased (P=0.005) by iPSC-EC treatment (0.577±0.118) as compared to the saline treatment group (0.436±0.040). This study suggests that iPSC-ECs may be useful in therapeutic strategies to ameliorate ischemic syndromes.
- © 2010 by American Heart Association, Inc.