Factors secreted by endothelial progenitor cells enhance neurorepair responses after cerebral ischemia in mice

PLoS One. 2013 Sep 4;8(9):e73244. doi: 10.1371/journal.pone.0073244. eCollection 2013.

Abstract

Cell therapy with endothelial progenitor cells (EPCs) has emerged as a promising strategy to regenerate the brain after stroke. Here, we aimed to investigate if treatment with EPCs or their secreted factors could potentiate angiogenesis and neurogenesis after permanent focal cerebral ischemia in a mouse model of ischemic stroke. BALB/C male mice were subjected to distal occlusion of the middle cerebral artery, and EPCs, cell-free conditioned media (CM) obtained from EPCs, or vehicle media were administered one day after ischemia. Magnetic resonance imaging (MRI) was performed at baseline to confirm that the lesions were similar between groups. Immunohistochemical and histological evaluation of the brain was performed to evaluate angio-neurogenesis and neurological outcome at two weeks. CM contained growth factors, such as VEGF, FGF-b and PDGF-bb. A significant increase in capillary density was noted in the peri-infarct areas of EPC- and CM-treated animals. Bielschowsky's staining revealed a significant increase in axonal rewiring in EPC-treated animals compared with shams, but not in CM-treated mice, in close proximity with DCX-positive migrating neuroblasts. At the functional level, post-ischemia forelimb strength was significantly improved in animals receiving EPCs or CM, but not in those receiving vehicle media. In conclusion, we demonstrate for the first time that the administration of EPC-secreted factors could become a safe and effective cell-free option to be considered in future therapeutic strategies for stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / pathology
  • Brain / pathology*
  • Brain / physiopathology*
  • Brain Ischemia / complications
  • Brain Ischemia / pathology
  • Brain Ischemia / physiopathology
  • Brain Ischemia / therapy*
  • Cell Movement
  • Cell- and Tissue-Based Therapy / methods*
  • Culture Media, Conditioned / metabolism
  • Doublecortin Protein
  • Endothelial Cells / cytology*
  • Magnetic Resonance Imaging
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Neovascularization, Physiologic
  • Stem Cells / cytology*
  • Stroke / complications
  • Treatment Outcome

Substances

  • Culture Media, Conditioned
  • Dcx protein, mouse
  • Doublecortin Protein

Grants and funding

AR is supported by the Miguel Servetprogramme (CP09/00265) from the Spanish Ministry of Health (Instituto de Salud Carlos III). LGB holds a Sara Borrell grant, and EMS holds a Rio Ortega grant from the Spanish Ministry of Health (Instituto de Salud Carlos III). This work has been supported by research grants from the Fondo de InvestigacionesSanitarias (PI10/00694), the Spanish stroke research network RENEVAS (RD06/0026/0010) and EUROSALUD programme (MICCIN). The research leading to these results has received funding from the European Union’s Seventh Framework Programme (FP7/2007–2013) under grant agreement numbers 201024 and 202213 (European Stroke Network). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.