通过超声将反义寡脱氧核苷酸体外转染至冠状动脉内皮细胞。

In vitro transfer of antisense oligodeoxynucleotides into coronary endothelial cells by ultrasound.

作者信息

Miura Shin-ichiro, Tachibana Katsuro, Okamoto Takahiro, Saku Keijiro

机构信息

Department of Cardiology, Fukuoka University School of Medicine, 7-45-1 Nanakuma, Jonan-Ku, Fukuoka 814-0180, Japan.

出版信息

Biochem Biophys Res Commun. 2002 Nov 8;298(4):587-90. doi: 10.1016/s0006-291x(02)02467-1.

DOI:10.1016/s0006-291x(02)02467-1

PMID:12408992

Abstract

Since antisense oligodeoxynucleotides (AS-ODNs) have been recognized as a new generation of putative therapeutic agents, we established a delivery technique that could transfect AS-ODNs, which are designed for endothelin type B receptor (ETB), into cultured human coronary endothelial cells (HCECs) by exposure to ultrasound in the presence of echo contrast microbubbles. Ultrasound offers several advantages such as being nontoxic, nonantigenic and providing rapid gene transfer. We standardized the optimal conditions, which consisted of 2 x 10(6) cells suspended in phosphate buffer with 900nM ODN, 50 microl of echo contrast microbubbles (Optison), and ultrasound exposure (1.0 W/cm(2), 10% duty cycle, and 10s duration). The percentage of transfected cells was 25.2+/-2.0% after ultrasound treatment. This is the first demonstration of the use of the ultrasound exposure technique in conjunction with microbubbles in HCECs.

摘要

由于反义寡脱氧核苷酸（AS - ODNs）已被公认为新一代潜在的治疗药物，我们建立了一种递送技术，该技术可在超声造影微泡存在的情况下，通过超声照射将针对内皮素B型受体（ETB）设计的AS - ODNs转染到培养的人冠状动脉内皮细胞（HCECs）中。超声具有多种优点，如无毒、无抗原性且能实现快速基因转移。我们对最佳条件进行了标准化，包括将2×10⁶个细胞悬浮于含有900nM ODN的磷酸盐缓冲液中，加入50微升超声造影微泡（Optison），并进行超声照射（1.0W/cm²，10%占空比，持续10秒）。超声处理后转染细胞的百分比为25.2±2.0%。这是首次证明在HCECs中使用超声照射技术结合微泡的情况。