title: Human platelet lysate improves human cord blood derived ECFC survival and vasculogenesis in three dimensional (3D) collagen matrices.
abstract: Human cord blood (CB) is enriched in circulating endothelial colony forming cells (ECFCs) that display high proliferative potential and in vivo vessel forming ability. Since diminished ECFC survival is known to dampen the vasculogenic response in vivo, we tested how long implanted ECFC survive and generate vessels in three-dimensional (3D) type I collagen matrices in vitro and in vivo. We hypothesized that human platelet lysate (HPL) would promote cell survival and enhance vasculogenesis in the 3D collagen matrices. We report that the percentage of ECFC co-cultured with HPL that were alive was significantly enhanced on days 1 and 3 post-matrix formation, compared to ECFC alone containing matrices. Also, co-culture of ECFC with HPL displayed significantly more vasculogenic activity compared to ECFC alone and expressed significantly more pro-survival molecules (pAkt, p-Bad and Bcl-xL) in the 3D collagen matrices in vitro. Treatment with Akt1 inhibitor (A-674563), Akt2 inhibitor (CCT128930) and Bcl-xL inhibitor (ABT-263/Navitoclax) significantly decreased the cell survival and vasculogenesis of ECFC co-cultured with or without HPL and implicated activation of the Akt1 pathway as the critical mediator of the HPL effect on ECFC in vitro. A significantly greater average vessel number and total vascular area of human CD31(+) vessels were present in implants containing ECFC and HPL, compared to the ECFC alone implants in vivo. We conclude that implantation of ECFC with HPL in vivo promotes vasculogenesis and augments blood vessel formation via diminishing apoptosis of the implanted ECFC.
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA; Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA; Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA. Electronic address: kimhyo@iu.edu.
authors.forename: Hyojin
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA; Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA. Electronic address: nprasain@iu.edu.
authors.forename: Nutan
authors.initials: N
authors.lastname: Prasain
authors.affiliation: Division of Hematology and Oncology, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA. Electronic address: svemula@iu.edu.
authors.forename: Sasidhar
authors.initials: S
authors.lastname: Vemula
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA; Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA. Electronic address: mferkow@iu.edu.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Ferkowicz
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA; Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA. Electronic address: myoshimo@iu.edu.
authors.forename: Momoko
authors.initials: M
authors.lastname: Yoshimoto
authors.affiliation: Weldon School of Biomedical Engineering, Purdue University, 206 S. Martin Jischke Drive, West Lafayette, IN 47907, USA. Electronic address: harbins@purdue.edu.
authors.forename: Sherry L
authors.initials: SL
authors.lastname: Voytik-Harbin
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA; Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA; Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, 1044 W. Walnut Street, Indianapolis, IN 46202, USA. Electronic address: myoder@iu.edu.
authors.forename: Mervin C
authors.initials: MC
authors.lastname: Yoder
mesh_terms: D000818:Animals; D017209:Apoptosis; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism*; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000737:chemistry*; D036101:Cord Blood Stem Cell Transplantation; D042783:Endothelial Cells / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D018919:Neovascularization, Physiologic / Q000502:physiology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1; D003094:Collagen; C494918:AKT1 protein, human; D051057:Proto-Oncogene Proteins c-akt
keywords: Apoptosis; Endothelial colony forming cells (ECFCs); Human platelet lysate (HPL); Three dimensional (3D) collagen matrix; Vasculogenesis