Improvement of Postnatal Neovascularization by Human Embryonic Stem Cell–Derived Endothelial-Like Cell Transplantation in a Mouse Model of Hindlimb Ischemia

Abstract

Background-We established an efficient preparation method to obtain endothelial-like cells (ECs) from human embryonic stem cells (hESCs) and tested whether these hESC-ECs would show therapeutic potential for treatment of hindlimb ischemia. Methods and Results-ECs differentiated from hESCs were obtained by mechanical isolation and cell sorting for von Willebrand factor. The isolated hESC-ECs maintained endothelial cell-specific characteristics such as endothelial marker expression and capillary formation. One day after surgical induction of hindlimb ischemia in athymic mice, hESC-ECs were injected intramuscularly into ischemic limbs. Four weeks after treatment, hESC-EC treatment significantly increased limb salvage (36%) compared with treatment with medium (0%). In addition, laser Doppler imaging showed that the ratio of blood perfusion (ischemic to normal limb) was increased significantly (PϽ0.01) by hESC-EC treatment (0.511Ϯ0.167) compared with medium injection (0.073Ϯ0.061). Capillary and arteriole densities were 658Ϯ190/mm 2 and 30Ϯ11/mm 2 in the hESC-EC group, respectively, whereas those in the medium group were 392Ϯ118/mm 2 and 16Ϯ8/mm

Genetic engineering of human stem cells for enhanced angiogenesis using biodegradable polymeric nanoparticles

“…Hindlimb ischemia was induced in a mouse model as previously described (6). Immediately after arterial dissection, cells (1.0 × 10 6 cells per injection) were suspended in 100 μL of hMSC growth medium and injected intramuscularly into two sites of the gracilis muscle in the medial thigh.…”

“…Stem cells are promising therapeutics for revascularization because of their capability of self-renewal, relative ease of isolation, and ability to migrate toward the ischemic tissues (4). Stem cells can contribute to angiogenesis directly, by participating in new vessel formation (5,6), or indirectly by secreting a broad spectrum of angiogenic and antiapoptotic factors (7,8). Furthermore, stem cells possess a homing capacity that allows them to migrate toward and engraft into the sites of ischemia or injury.…”

An electrochemical functional assay for the sensing of nitric oxide release induced by angiogenic factors

“…http://bmbreports.org Electrochemical measurements of ANG induced NO release were repeated using stem-cell derived endothelial cells, and similar responses were recorded (28). Differentiation of these stem cells into endothelial cells is usually assessed by immunostaining with anti-Von Willebrand factor (43), but use of NO release after addition of angiogenic factors could be used as a possible alternative criterion for differentiation. ANG has been reported to have 4 different mechanisms of action (44):…”

Engineering Strategies for the Formation of Embryoid Bodies from Human Pluripotent Stem Cells

“…formed in suspension culture for enrichment toward specific lineages [35,36]. The efficiency of these methods to enrich EBs for target lineage specifications is only moderate, and the involvement of labor-intensive procedures prevents scale-up.…”