Functional and Transcriptional Characterization of Human Embryonic Stem Cell-Derived Endothelial Cells for Treatment of Myocardial Infarction

Abstract

BackgroundDifferentiation of human embryonic stem cells into endothelial cells (hESC-ECs) has the potential to provide an unlimited source of cells for novel transplantation therapies of ischemic diseases by supporting angiogenesis and vasculogenesis. However, the endothelial differentiation efficiency of the conventional embryoid body (EB) method is low while the 2-dimensional method of co-culturing with mouse embryonic fibroblasts (MEFs) require animal product, both of which can limit the future clinical application of hESC-ECs. Moreover, to fully understand the beneficial effects of stem cell therapy, investigators must be able to track the functional biology and physiology of transplanted cells in living subjects over time.MethodologyIn this study, we developed an extracellular matrix (ECM) culture system for increasing endothelial differentiation and free from contaminating animal cells. We investigated the transcriptional changes that occur during endothelial differentiation of hESCs using whole genome microarray, and compared to human umbilical vein endothelial cells (HUVECs). We also showed functional vascular formation by hESC-ECs in a mouse dorsal window model. Moreover, our study is the first so far to transplant hESC-ECs in a myocardial infarction model and monitor cell fate using molecular imaging methods.ConclusionTaken together, we report a more efficient method for derivation of hESC-ECs that express appropriate patterns of endothelial genes, form functional vessels in vivo, and improve cardiac function. These studies suggest that hESC-ECs may provide a novel therapy for ischemic heart disease in the future.

Preclinical Derivation and Imaging of Autologously Transplanted Canine Induced Pluripotent Stem Cells

“…A left thoracotomy was performed followed by ligation of the left anterior descending artery for 30 min followed by reperfusion as described previously (15). After 30 min, 1 ϫ 10 6 ciPSC-ECs stably expressing Fluc-RFP-HSVttk were injected intramyocardially into three sites near the periinfarct zone at 20 ml of total volume (n ϭ 8).…”

“…Living Image 4.0 (Caliper Life Sciences) was used to quantify signal intensity as described previously (15,16).…”

“…Fractional shortening was acquired using standard M-mode image acquisitions of left ventricular short axis images as described previously (15,16).…”

“…Model-Previous studies have demonstrated potential of iPSCand ESC-derived endothelial cells to improve systolic function following myocardial infarction through revascularization (15,16). Hence, we next delivered ciPSC-ECs stably transduced with TF reporter gene into the border zone of infarcted murine hearts.…”

Human embryonic stem cells-derived endothelial cell therapy facilitates kidney regeneration by stimulating renal resident stem cell proliferation in acute kidney injury

“…Human ESC-ECs were obtained and cultured as previously described [14,15], and were transfected with a self-inactivating lentiviral vector, constructed with a ubiquitin promoter-driven firefly luciferase and an enhanced green fluorescence protein (Fluc-eGFP) double fusion reporter gene [14,22]. The transfected hESC-ECs were maintained in EGM-2 medium (Lonza, Basel, Switzerland) under standard cell culture conditions (37°C, 95% O 2 /5% CO 2 ) for 3 d prior to experimental use.…”