Efficient gene delivery to primary human retinal pigment epithelial cells: The innate and acquired properties of vectors

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• 作者：Nooshin Tasharrofi^a ; ^b ; ^{n_tasharrofi@yahoo.com} ; ^{atyabifa@tums.ac.ir} ; Fatemeh Kouhkan^c ; ^{f.kouhkan@stemcellstech.com} ; Masoud Soleimani^d ; ^{soleim_m@modares.ac.ir} ; Zahra-Soheila Soheili^e ; ^{soheili@nigeb.ac.ir} ; Fatemeh Atyabi^a ; ^b ; ^f ; Hamid Akbari Javar^b ; ^{akbarijo@tums.ac.ir} ; Farid Abedin Dorkoosh^b ; ^g ; ^{dorkoosh@tums.ac.ir}
• 关键词：PLGA/PEI nanoparticles ; Primary human RPE cells ; Transfection ; Vitreous ; Protein adsorption
• 刊名：International Journal of Pharmaceutics
• 出版年：2017
• 出版时间：25 February 2017
• 年：2017
• 卷：518
• 期：1-2
• 页码：66-79
• 全文大小：3530 K
• 卷排序：518

文摘

The purpose of this study is designing non-viral gene delivery vectors for transfection of the primary human retinal pigment epithelial cells (RPE). In the design process of gene delivery vectors, considering physicochemical properties of vectors alone does not seem to be enough since they interact with constituents of the surrounding environment and hence gain new characteristics. Moreover, due to these interactions, their cargo can be released untimely or undergo degradation before reaching to the target cells. Further, the characteristics of cells itself can also influence the transfection efficacy. For example, the non-dividing property of RPE cells can impede the transfection efficiency which in most studies was ignored by using immortal cell lines. In this study, vectors with different characteristics differing in mixing orders of pDNA, PEI polymer, and PLGA/PEI or PLGA nanoparticles were prepared and characterized. Then, their characteristics and efficacy in gene delivery to RPE cells in the presence of vitreous or fetal bovine serum (FBS) were evaluated. All formulations showed no cytotoxicity and were able to protect pDNA from premature release and degradation in extracellular media. Also, the adsorption of vitreous or serum proteins onto the surface of vectors changed their properties and hence cellular uptake and transfection efficacy.