A flow cytometric approach to analyzing mature and progenitor endothelial cells following traumatic brain injury

Poincyane Assis-Nascimento, Oliver Umland, Maria L. Cepero, Daniel J Liebl

Research output: Contribution to journalArticle

6 Citations (Scopus)

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. New method: 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. Comparison with existing 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.

Original languageEnglish (US)
Pages (from-to)57-67
Number of pages11
JournalJournal of Neuroscience Methods
Volume263
DOIs
StatePublished - Apr 1 2016

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Flow Cytometry
Endothelial Cells
Wounds and Injuries
Blood-Brain Barrier
Leukocytes
CD31 Antigens
Vascular Endothelial Growth Factor Receptor-2
Blood Vessels
Cell Count
Fluorescence
Western Blotting
Traumatic Brain Injury
Endothelial Progenitor Cells
Brain
Proteins

Keywords

  • Blood
  • Endothelial cells
  • Endothelial progenitor cells
  • Flow cytometry
  • Traumatic brain injury
  • Vessels

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

A flow cytometric approach to analyzing mature and progenitor endothelial cells following traumatic brain injury. / Assis-Nascimento, Poincyane; Umland, Oliver; Cepero, Maria L.; Liebl, Daniel J.

In: Journal of Neuroscience Methods, Vol. 263, 01.04.2016, p. 57-67.

Research output: Contribution to journalArticle

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