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使用核转染技术高效转染原代兔泪腺腺泡细胞。

Use of nucleofection to efficiently transfect primary rabbit lacrimal gland acinar cells.

作者信息

Contreras Janette, Hsueh Pang-Yu, Pei Hua, Hamm-Alvarez Sarah F

机构信息

Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, Los Angeles, CA, 90033, USA.

出版信息

Cytotechnology. 2012 Mar;64(2):149-56. doi: 10.1007/s10616-011-9404-3. Epub 2011 Dec 3.

Abstract

Lacrimal gland acinar cells are an important cell type to study due to their role in production and release of tear proteins, a function essential for ocular surface integrity and normal visual acuity. However, mechanistic studies are often limited by problems with transfection using either plasmid DNA or siRNA. Although various gene delivery methods are available, many have been unproductive due to consistently low transfection efficiencies. We have developed a method using nucleofection that can result in 50% transfection efficiency and 60% knockdown efficiency for plasmid DNA and siRNA, respectively. These results are vastly improved relative to previous studies, demonstrating that nucleofection offers an efficient transfection technique for primary lacrimal gland acinar cells.

摘要

泪腺腺泡细胞是一种重要的细胞类型,因其在泪液蛋白的产生和释放中发挥作用,而泪液蛋白对于眼表完整性和正常视力至关重要。然而,使用质粒DNA或小干扰RNA(siRNA)进行转染时存在的问题常常限制了机制研究。尽管有多种基因递送方法可供使用,但由于转染效率一直较低,许多方法都未取得成效。我们开发了一种使用核转染的方法,该方法分别可使质粒DNA和siRNA的转染效率达到50%,敲低效率达到60%。相对于之前的研究,这些结果有了极大的改善,表明核转染为原代泪腺腺泡细胞提供了一种高效的转染技术。

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