Improvement of postnatal neovascularization by human embryonic stem cell derived endothelial-like cell transplantation in a mouse model of hindlimb ischemia.
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.


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.
Cho
Seung-Woo
SW
Department of Bioengineering, Hanyang University, Seoul, Korea.
Moon
Sung-Hwan
SH
Lee
Soo-Hong
SH
Kang
Sun-Woong
SW
Kim
Jumi
J
Lim
Jae Min
JM
Kim
Hyo-Soo
HS
Kim
Byung-Soo
BS
Chung
Hyung-Min
HM
D000818:Animals; D000831:Animals, Newborn; D002460:Cell Line; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000637:transplantation*; D042783:Endothelial Cells / Q000637:transplantation*; D005260:Female; D006614:Hindlimb / Q000098:blood supply* / Q000601:surgery; D006801:Humans; D007511:Ischemia / Q000473:pathology / Q000503:physiopathology* / Q000601:surgery*; D051379:Mice; D008819:Mice, Nude; D018919:Neovascularization, Physiologic / Q000502:physiology*; D033581:Stem Cell Transplantation / Q000379:methods