Transcriptomic analysis of the harvested endothelial cells in a swine model of mechanical thrombectomy

Neuroradiology. 2018 Jul;60(7):759-768. doi: 10.1007/s00234-018-2033-1. Epub 2018 May 14.

Abstract

Purpose: In mechanical thrombectomy (MT) for ischemic stroke, endothelial cells (ECs) from intracranial blood vessels adhere to the stent retriever device and can be harvested. However, understanding the molecular biology and the role of the endothelium in different pathological conditions remains insufficient. The purpose of the study was to characterize and analyze the molecular aspect of harvested ECs using cell culture and transcriptomic techniques in an MT swine model relevant to clinical ischemic stroke.

Methods: In swine, preformed thrombi were injected into the external carotid and subclavian arteries to occlude their branches. MT was performed according to clinical routine. The stent retriever device and thrombus were treated with cell dissociation buffer. The resulting cell suspension was analyzed by immunohistochemistry and was cultured. Cultured cells were analyzed using single-cell RNA sequencing (scRNA-seq) after fluorescence-activated cell sorting (FACS).

Results: A total number of 37 samples were obtained containing CD31-positive cells. Cell culture was successful in 90% of samples, and the cells expressed multiple typical EC protein markers. Eighty-nine percent of the sorted cells yielded high-quality transcriptomes, and single-cell transcriptomes from cultured cells showed that they expressed typical endothelial gene patterns. Gene expression analysis of ECs from an occluded artery did not show distinctive clustering into subtypes.

Conclusion: ECs harvested during MT can be cultured and analyzed using single-cell transcriptomic techniques. This analysis can be implemented in clinical practice to study the EC gene expression of comorbidities, such as hypertension, diabetes mellitus, and metabolic syndrome, in patients suffering from acute ischemic stroke.

Keywords: Cell culture; Endothelial cells; Thrombectomy; Transcriptomic analysis.

MeSH terms

  • Animals
  • Cells, Cultured
  • Cerebral Angiography
  • Endothelial Cells / pathology*
  • Flow Cytometry
  • Gene Expression Profiling / methods*
  • Immunohistochemistry
  • Microscopy, Electron, Scanning
  • Principal Component Analysis
  • RNA / genetics*
  • Stroke / genetics*
  • Stroke / pathology
  • Swine
  • Thrombectomy / methods*

Substances

  • RNA