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

• Published in: Biotechnology reports (Amsterdam, Netherlands) Vol. 7; no. C; pp. 157 - 165
• Main Authors: Sun, Zhengda, Lawson, Devon A., Sinclair, Elizabeth, Wang, Chih-Yang, Lai, Ming-Derg, Hetts, Steven W., Higashida, Randall T., Dowd, Christopher F., Halbach, Van V., Werb, Zena, Su, Hua, Cooke, Daniel L.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2015; Elsevier
• Subjects: Gene expression of artery endothelial cells; Single cell quantitative RT-PCR; Targeted endothelial cell sampling; Targeted endothelial cell sampling; Single cell quantitative RT-PCR; Gene expression of artery endothelial cells
• ISSN: 2215-017X; 2215-017X
• DOI: 10.1016/j.btre.2015.07.001

[Display omitted] •The combination of guide wire sampling, FACS and high throughput microfluidic single-cell quantitative RT-PCR, is an effective strategy for analyzing molecular changes of ECs in vascular lesions.•Although heterogeneous, the ECs in normal iliac artery fall into two classes. 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. 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. 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.