Generation of an anti-angiogenic endothelial progenitor cell line via endostatin gene transfer.

Molecular Medicine Reports
Jing AiKe Yao

Abstract

The viability of endothelial progenitor cells (EPCs) as a therapeutic treatment for neovascularization (NV) was subject to investigation in the present study. Furthermore, endostatin has previously been demonstrated to be an inhibitor of angiogenesis and a suppressant of vascular leakage. The aim of the present study was to generate transgenic EPCs with anti‑angiogenic effects for the treatment of ocular NV. EPCs were obtained from rat peripheral blood samples and then verified. A lentiviral‑endostatin‑green fluorescent protein recombinant construct was generated and used to infect EPCs. Transfected cells were then subjected to puromycin selection. Reverse transcription‑quantitative polymerase chain reaction and a western blot assay were then applied in order to determine both the endostatin mRNA and protein expression levels, respectively. In addition, vascular endothelial growth factor (VEGF) expression levels were also detected in order to observe the anti‑angiogenic effect of the endostatin‑transfected EPCs. Following puromycin (1 µg/ml) selection for 4 days, a stable endostatin‑transfected EPC line was generated. In this stable endostatin-transfected EPC line, the expression levels of endostatin increased; whereas the expr...Continue Reading

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Citations

Aug 17, 2020·International Journal of Molecular Sciences·Francesca TinelliLaura Gatti

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Methods Mentioned

BETA
transfection
fluorescence microscopy
flow cytometry
electrophoresis
protein assay

Software Mentioned

CellQuest Pro
ClonExpress
SPSS
Pro Analyzer
Gel

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