title: Role of VEGF-A in angiogenesis promoted by umbilical cord-derived mesenchymal stromal/stem cells: in vitro study.
abstract: BACKGROUND
  Mesenchymal stromal/stem cells derived from human umbilical cord (UC-MSCs) uniquely combine properties of embryonic and postnatal MSCs and may be the most acceptable, safe, and effective source for allogeneic cell therapy e.g. for therapeutic angiogenesis. In this report we describe pro-angiogenic properties of UC-MSCs as manifested in vitro.
  
  
  METHODS
  UC-MSCs were isolated from human Wharton's jelly by enzymatic digestion. Presence of soluble forms of VEGF-A in UC-MSC-conditioned media was measured by ELISA. Effects of the conditioned media on human umbilical vein-derived endothelial EA.hy926 cells proliferation were measured by MTT-assay; changes in cell motility and directed migration were assessed by scratch wound healing and transwell chamber migration assays. Angiogenesis was modeled in vitro as tube formation on basement membrane matrix. Progressive differentiation of MSCs to endothelioid progeny was assessed by CD31 immunostaining.
  
  
  RESULTS
  Although no detectable quantities of soluble VEGF-A were produced by UC-MSCs, the culture medium, conditioned by the UC-MSCs, effectively stimulated proliferation, motility, and directed migration of EA.hy926 cells. In 2D culture, UC-MSCs were able to acquire CD31(+) endothelial cell-like phenotype when stimulated by EA.hy926-conditioned media supplemented with VEGF-A165. UC-MSCs were capable of forming unstable 2D tubular networks either by themselves or in combinations with EA.hy926 cells. Active spontaneous sprouting from cell clusters, resulting from disassembling of such networks, was observed only in the mixed cultures, not in pure UC-MSC cultures. In 3D mode of sprouting experimentation, structural support of newly formed capillary-like structures was provided by UC-MSCs that acquired the CD31(+) phenotype in the absence of exogenous VEGF-A.
  
  
  CONCLUSIONS
  These data suggest that a VEGF-A-independent paracrine mechanism and at least partially VEGF-A-independent differentiation mechanism are involved in the pro-angiogenic activity of UC-MSCs.
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia.|Scientific Research Institute of Human Morphology, 3 Tsurupa Street, Moscow, 117418, Russia.
authors.forename: Irina
authors.initials: I
authors.lastname: Arutyunyan
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia. tfat@yandex.ru.|Pirogov Russian National Research Medical University, Ministry of Healthcare of the Russian Federation, 1 Ostrovitianov Street, Moscow, 117997, Russia. tfat@yandex.ru.|Laboratory of Regenerative Medicine, Research Center for Obstetrics, Gynecology and Perinatology, 4 Oparin Street, Moscow, 117997, Russia. tfat@yandex.ru.
authors.forename: Timur
authors.initials: T
authors.lastname: Fatkhudinov
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia.|Scientific Research Institute of Human Morphology, 3 Tsurupa Street, Moscow, 117418, Russia.
authors.forename: Evgeniya
authors.initials: E
authors.lastname: Kananykhina
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia.|Pirogov Russian National Research Medical University, Ministry of Healthcare of the Russian Federation, 1 Ostrovitianov Street, Moscow, 117997, Russia.
authors.forename: Natalia
authors.initials: N
authors.lastname: Usman
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia.|Scientific Research Institute of Human Morphology, 3 Tsurupa Street, Moscow, 117418, Russia.
authors.forename: Andrey
authors.initials: A
authors.lastname: Elchaninov
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia.|Pirogov Russian National Research Medical University, Ministry of Healthcare of the Russian Federation, 1 Ostrovitianov Street, Moscow, 117997, Russia.
authors.forename: Andrey
authors.initials: A
authors.lastname: Makarov
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia.
authors.forename: Galina
authors.initials: G
authors.lastname: Bolshakova
authors.affiliation: Research Center of Medical Genetics, 1 Moskvorechie Street, Moscow, 115478, Russia.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Goldshtein
authors.affiliation: Research Center for Obstetrics, Gynecology and Perinatology of Ministry of Healthcare of the Russian Federation, 4 Oparina Street, Moscow, 117997, Russia.
authors.forename: Gennady
authors.initials: G
authors.lastname: Sukhikh
mesh_terms: D002454:Cell Differentiation; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D017077:Culture Media, Conditioned; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D018919:Neovascularization, Physiologic; D014470:Umbilical Cord / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000502:physiology*; D059631:Wharton Jelly / Q000166:cytology
chemical_list: D017077:Culture Media, Conditioned; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: Angiogenesis inducing agents; CD31 antigen; Cell migration assays; Endothelial cells; Extracellular matrix; In vitro techniques; Mesenchymal stromal cells; Multipotent; Vascular endothelial growth factor-A; Wharton jelly