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Biotechnol Rep (Amst). 2015 Sep;7:157-165. Epub 2015 Aug 1.

Endovascular biopsy: Strategy for analyzing gene expression profiles of individual endothelial cells obtained from human vessels.

Author information

1
Division of Neurointerventional Radiology, Department of Radiology and Biomedical Imaging, University of California, San Francisco, CA, USA.
2
Department of Anatomy, University of California, San Francisco, CA, USA.
3
Division of Experimental Medicine, Department of Medicine, University of California, San Francisco, CA, USA.
4
Department of Anatomy, University of California, San Francisco, CA, USA; Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
5
Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
6
Center for Cerebrovascular Research, Department of Anesthesia and Perioperative Care, University of California, San Francisco, CA, USA.

Abstract

PURPOSE:

To develop a strategy of achieving targeted collection of endothelial cells (ECs) by endovascular methods and analyzing the gene expression profiles of collected single ECs.

METHODS AND RESULTS:

134 ECs and 37 leukocytes were collected from four patients' intra-iliac artery endovascular guide wires by fluorescence activated cell sorting (FACS) and analyzed by single-cell quantitative RT-PCR for expression profile of 48 genes. Compared to CD45+ leukocytes, the ECs expressed higher levels (p < 0.05) of EC surface markers used on FACS and other EC related genes. The gene expression profile showed that these isolated ECs fell into two clusters, A and B, that differentially expressed 19 genes related to angiogenesis, inflammation and extracellular matrix remodeling, with cluster B ECs have demonstrating similarities to senescent or aging ECs.

CONCLUSION:

Combination of endovascular device sampling, FACS and single-cell quantitative RT-PCR is a feasible method for analyzing EC gene expression profile in vascular lesions.

KEYWORDS:

Gene expression of artery endothelial cells; Single cell quantitative RT-PCR; Targeted endothelial cell sampling

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