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

Neuroradiology
Nasren JaffStaffan Holmin

Abstract

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. 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). 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 tran...Continue Reading

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Citations

Jan 19, 2019·Neuroradiology·Rüdiger von Kummer
Aug 11, 2018·Molecular Therapy. Methods & Clinical Development·Aisha A AlJanahiCynthia E Dunbar
Jul 13, 2021·Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism·Benjamin C ShawAnn M Stowe

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Methods Mentioned

BETA
biopsy
RNA-seq
dissection
fluorescence-activated cell sorting
FACS
scanning electron microscopy
X-ray
scRNA-seq
PCA

Software Mentioned

Bioconductor for R
DESeq2
ccRemover
QualiMap
FastQC

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