title: A flow cytometric approach to analyzing mature and progenitor endothelial cells following traumatic brain injury.
abstract: BACKGROUND
  Traumatic brain injury (TBI) continues to be a major source of death and disability worldwide, and one of the earliest and most profound deficits comes from vascular damage and breakdown of the blood-brain barrier (BBB). Cerebral vascular endothelial cells (cvECs) and endothelial progenitor cells (EPCs) have been shown to play essential roles in vessel repair and BBB stability, although their individual contributions remain poorly defined.
  
  
  METHODS
  We employ TruCount beads with flow cytometry to precisely quantify cvECs, EPCs, and peripheral leukocytes in the murine cortex after controlled cortical impact (CCI) injury.
  
  
  RESULTS
  We found a significant reduction in the number of cvECs at 3 days post-injury (dpi), whereas the EPCs and invading peripheral leukocytes were significantly increased compared with sham controls. Proliferation studies demonstrate that both cvECs and EPCs are undergoing cell expansion in the first week post-injury. Furthermore, analysis of protein expression using mean fluorescence intensity found increases in PECAM-1, VEGFR-2, and VE-Cadherin expression per cell at 3 dpi, which is consistent with western blot analysis.
  
  
  METHODS
  Classic methods of cell analysis, such as histological cell counts, in the traumatic injured brain are labor intensive, time-consuming, and potentially biased; whereas flow cytometry provides an efficient, non-biased approach to simultaneously quantify multiple cell types. However, conventional flow cytometry that employs capped events can provide misleading results in CNS injured tissues.
  
  
  CONCLUSIONS
  We demonstrate that TruCount quantification using flow cytometry is a powerful tool for quantifying mature and progenitor endothelial cell changes after TBI.
authors: The Miami Project to Cure Paralysis, the Department of Neurosurgery, the University of Miami, Miller School of Medicine, Miami, FL, USA.
authors: Poincyane
authors: P
authors: Assis-Nascimento
authors: Diabetes Research Institute, the University of Miami, Miller School of Medicine, Miami, FL, USA.
authors: Oliver
authors: O
authors: Umland
authors: The Miami Project to Cure Paralysis, the Department of Neurosurgery, the University of Miami, Miller School of Medicine, Miami, FL, USA.
authors: Maria L
authors: ML
authors: Cepero
authors: The Miami Project to Cure Paralysis, the Department of Neurosurgery, the University of Miami, Miller School of Medicine, Miami, FL, USA. Electronic address: dliebl@miami.edu.
authors: Daniel J
authors: DJ
authors: Liebl
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D000070642:Brain Injuries, Traumatic / Q000473:pathology*; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D049109:Cell Proliferation / Q000502:physiology*; D004195:Disease Models, Animal; D066026:Endothelial Progenitor Cells / Q000473:pathology*; D005434:Flow Cytometry; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D010671:Phenylurea Compounds / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D015815:Cell Adhesion Molecules; D008164:Luminescent Proteins; D010671:Phenylurea Compounds; D000099152:Cadherin 5; C013065:cell aggregation factors; C032766:ethylene diurea
keywords: Blood; Endothelial cells; Endothelial progenitor cells; Flow cytometry; Traumatic brain injury; Vessels