title: Spatially oriented, temporally sequential smooth muscle cell-endothelial progenitor cell bi-level cell sheet neovascularizes ischemic myocardium.
abstract: BACKGROUND
  Endothelial progenitor cells (EPCs) possess robust therapeutic angiogenic potential, yet may be limited in the capacity to develop into fully mature vasculature. This problem might be exacerbated by the absence of a neovascular foundation, namely pericytes, with simple EPC injection. We hypothesized that coculturing EPCs with smooth muscle cells (SMCs), components of the surrounding vascular wall, in a cell sheet will mimic the native spatial orientation and interaction between EPCs and SMCs to create a supratherapeutic angiogenic construct in a model of ischemic cardiomyopathy.
  
  
  RESULTS
  Primary EPCs and SMCs were isolated from Wistar rats. Confluent SMCs topped with confluent EPCs were spontaneously detached from the Upcell dish to create an SMC-EPC bi-level cell sheet. A rodent ischemic cardiomyopathy model was created by ligating the left anterior descending coronary artery. Rats were then immediately divided into 3 groups: cell-sheet transplantation (n=14), cell injection (n=12), and no treatment (n=13). Cocultured EPCs and SMCs stimulated an abundant release of multiple cytokines in vitro. Increased capillary density and improved blood perfusion in the borderzone elucidated the significant in vivo angiogenic potential of this technology. Most interestingly, however, cell fate-tracking experiments demonstrated that the cell-sheet EPCs and SMCs directly migrated into the myocardium and differentiated into elements of newly formed functional vasculature. The robust angiogenic effect of this cell sheet translated to enhanced ventricular function as demonstrated by echocardiography.
  
  
  CONCLUSIONS
  Spatially arranged EPC-SMC bi-level cell-sheet technology facilitated the natural interaction between EPCs and SMCs, thereby creating structurally mature, functional microvasculature in a rodent ischemic cardiomyopathy model, leading to improved myocardial function.
authors.affiliation: Division of Cardiovascular Surgery, Department of Surgery, University of Pennsylvania School of Medicine, Philadelphia, PA (Y.S., J.E.C., J.W.M., P.A., P.F.H., E.C.Y., A.S.F., A.T., J.P., Y.J.W.); and Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Osaka, Japan (S.M., Y.S.).
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Shudo
authors.forename: Jeffrey E
authors.initials: JE
authors.lastname: Cohen
authors.forename: John W
authors.initials: JW
authors.lastname: Macarthur
authors.forename: Pavan
authors.initials: P
authors.lastname: Atluri
authors.forename: Philip F
authors.initials: PF
authors.lastname: Hsiao
authors.forename: Elaine C
authors.initials: EC
authors.lastname: Yang
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Fairman
authors.forename: Alen
authors.initials: A
authors.lastname: Trubelja
authors.forename: Jay
authors.initials: J
authors.lastname: Patel
authors.forename: Shigeru
authors.initials: S
authors.lastname: Miyagawa
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Sawa
authors.forename: Y Joseph
authors.initials: YJ
authors.lastname: Woo
mesh_terms: D000818:Animals; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000473:pathology / Q000502:physiology*; D005260:Female; D006801:Humans; D008297:Male; D017202:Myocardial Ischemia / Q000473:pathology*; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D009389:Neovascularization, Pathologic / Q000473:pathology*; D051381:Rats; D017208:Rats, Wistar; D013234:Stem Cells / Q000473:pathology / Q000502:physiology*; D013997:Time Factors
keywords: angiogenesis; cardiovascular diseases; cells; endothelium; heart failure; tissue
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