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 (P<0.01) by hESC-EC treatment (0.511+/-0.167) compared with medium injection (0.073+/-0.061). Capillary and arteriole densities were 658+/-190/mm2 and 30+/-11/mm2 in the hESC-EC group, respectively, whereas those in the medium group were 392+/-118/mm2 and 16+/-8/mm2, respectively (P<0.01). Reverse-transcription polymerase chain reaction with human-specific primers revealed mRNA expression of human endothelial markers and human angiogenic factors in ischemic mouse tissues. The transplanted hESC-ECs were localized as capillaries near muscle tissues in ischemic regions or incorporated in the vessels between muscle tissues, as confirmed by human nuclear antigen staining with platelet/endothelial cell adhesion molecule or von Willebrand factor. CONCLUSIONS: This study demonstrates that hESC-EC transplantation improves blood perfusion and limb salvage by facilitating postnatal neovascularization in a mouse model of hindlimb ischemia. Thus, hESC-ECs might be useful as an alternative cell source for angiogenic therapy.
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 (P<0.01) by hESC-EC treatment (0.511+/-0.167) compared with medium injection (0.073+/-0.061). Capillary and arteriole densities were 658+/-190/mm2 and 30+/-11/mm2 in the hESC-EC group, respectively, whereas those in the medium group were 392+/-118/mm2 and 16+/-8/mm2, respectively (P<0.01). Reverse-transcription polymerase chain reaction with human-specific primers revealed mRNA expression of human endothelial markers and human angiogenic factors in ischemicmouse tissues. The transplanted hESC-ECs were localized as capillaries near muscle tissues in ischemic regions or incorporated in the vessels between muscle tissues, as confirmed by human nuclear antigen staining with platelet/endothelial cell adhesion molecule or von Willebrand factor. CONCLUSIONS: This study demonstrates that hESC-EC transplantation improves blood perfusion and limb salvage by facilitating postnatal neovascularization in a mouse model of hindlimb ischemia. Thus, hESC-ECs might be useful as an alternative cell source for angiogenic therapy.
Authors: Mingxia Gu; Nicholas M Mordwinkin; Nigel G Kooreman; Jaecheol Lee; Haodi Wu; Shijun Hu; Jared M Churko; Sebastian Diecke; Paul W Burridge; Chunjiang He; Frances E Barron; Sang-Ging Ong; Joseph D Gold; Joseph C Wu Journal: Eur Heart J Date: 2014-11-02 Impact factor: 29.983
Authors: Jung Ki Yoo; Jumi Kim; Seong-Jun Choi; Hye Min Noh; Young Do Kwon; Hanna Yoo; Hyo Seon Yi; Hyung Min Chung; Jin Kyeoung Kim Journal: Stem Cells Dev Date: 2012-02-07 Impact factor: 3.272
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Authors: Marilyn B Nourse; Daniel E Halpin; Marta Scatena; Derek J Mortisen; Nathaniel L Tulloch; Kip D Hauch; Beverly Torok-Storb; Buddy D Ratner; Lil Pabon; Charles E Murry Journal: Arterioscler Thromb Vasc Biol Date: 2009-10-29 Impact factor: 8.311
Authors: Jin Yu; Ngan F Huang; Kitchener D Wilson; Jeffrey B Velotta; Mei Huang; Zongjin Li; Andrew Lee; Robert C Robbins; John P Cooke; Joseph C Wu Journal: PLoS One Date: 2009-09-15 Impact factor: 3.240