title: Endothelial cells derived from human iPSCS increase capillary density and improve perfusion in a mouse model of peripheral arterial disease.
abstract: OBJECTIVE
  Stem cell therapy for angiogenesis and vascular regeneration has been investigated using adult or embryonic stem cells. In the present study, we investigated the potential of endothelial cells (ECs) derived from human induced pluripotent stem cells (hiPSCs) to promote the perfusion of ischemic tissue in a murine model of peripheral arterial disease.
  
  
  RESULTS
  Endothelial differentiation was initiated by culturing hiPSCs for 14 days in differentiation media supplemented with BMP-4 and vascular endothelial growth factor. The hiPSC-ECs exhibited endothelial characteristics by forming capillary-like structures in matrigel and incorporating acetylated-LDL. They stained positively for EC markers such as KDR, CD31, CD144, and eNOS. In vitro exposure of hiPSC-ECs to hypoxia resulted in increased expression of various angiogenic related cytokines and growth factors. hiPSC-ECs were stably transduced with a double fusion construct encoded by the ubiquitin promoter, firefly luciferase for bioluminescence imaging and green fluorescence protein for fluorescent detection. The hiPSC-ECs (5×10(5)) were delivered by intramuscular injection into the ischemic hindlimb of SCID mice at day 0 and again on day 7 after femoral artery ligation (n=8). Bioluminescence imaging showed that hiPSC-ECs survived in the ischemic limb for at least 2 weeks. In addition, laser Doppler imaging showed that the ratio of blood perfusion was increased by hiPSC-EC treatment by comparison to the saline-treated group (0.58±0.12 versus 0.44±0.04; P=0.005). The total number of capillaries in the ischemic limb of mice receiving hiPSC-EC injections was greater than those in the saline-treated group (1284±155 versus 797±206 capillaries/mm(2)) (P<0.002).
  
  
  CONCLUSIONS
  This study is a first step toward development of a regenerative strategy for peripheral arterial disease based on the use of ECs derived from hiPSCs.
authors.affiliation: Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Abdul Jalil
authors.initials: AJ
authors.lastname: Rufaihah
authors.forename: Ngan F
authors.initials: NF
authors.lastname: Huang
authors.forename: Sina
authors.initials: S
authors.lastname: Jamé
authors.forename: Jerry C
authors.initials: JC
authors.lastname: Lee
authors.forename: Ha N
authors.initials: HN
authors.lastname: Nguyen
authors.forename: Blake
authors.initials: B
authors.lastname: Byers
authors.forename: Abhijit
authors.initials: A
authors.lastname: De
authors.forename: Janet
authors.initials: J
authors.lastname: Okogbaa
authors.forename: Mark
authors.initials: M
authors.lastname: Rollins
authors.forename: Renee
authors.initials: R
authors.lastname: Reijo-Pera
authors.forename: Sanjiv S
authors.initials: SS
authors.lastname: Gambhir
authors.forename: John P
authors.initials: JP
authors.lastname: Cooke
mesh_terms: D000818:Animals; D002196:Capillaries / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D006614:Hindlimb / Q000098:blood supply; D006801:Humans; D007511:Ischemia / Q000628:therapy; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D018919:Neovascularization, Physiologic / Q000502:physiology; D058729:Peripheral Arterial Disease / Q000503:physiopathology* / Q000628:therapy*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D033581:Stem Cell Transplantation / Q000379:methods; D016896:Treatment Outcome
chemical_list: D016207:Cytokines