The relationship between terminal functionalization and molecular weight of a gene delivery polymer and transfection efficacy in mammary epithelial 2-D cultures and 3-D organotypic cultures.
The relationship between terminal functionalization and molecular weight of a gene delivery polymer and transfection efficacy in mammary epithelial 2-D cultures and 3-D organotypic cultures.
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DOI：
10.1016/j.biomaterials.2010.07.023
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发表时间：
2010-11
期刊：
影响因子：
14
通讯作者：
Green, Jordan J.
中科院分区：
文献类型：
--
作者：
Bhise, Nupura S.;Gray, Ryan S.;Sunshine, Joel C.;Htet, Soe;Ewald, Andrew J.;Green, Jordan J.
非病毒基因递送载体被开发用于有效地将基因转移到难以转染的小鼠乳腺上皮细胞。在传统的2-D单层和3-D器官型培养物中测试了相同基础聚（β-氨基酯），聚（1，4-丁二醇二丙烯酸酯-共-5-氨基-1-戊醇）的10个改性版本。聚合物与编码增强型绿色荧光蛋白的质粒DNA自组装形成用于转染细胞的纳米颗粒（~100 nm）。通过改变合成和制造条件来调整纳米颗粒转染功效，并使用共聚焦显微镜和流式细胞术分析转染功效。表现最好的聚合物纳米颗粒在2-D培养物中转染了57±6%的细胞，在3-D培养物中转染了6±1%的细胞。对聚合物封端分子的小修饰和聚合物分子量的调整可以显著增强转染功效高达6倍或完全消除功效。领先聚合物的功效在2-D中比商业转染剂FuGENE® HD高13倍，在3-D中高2倍。这些非病毒纳米颗粒可用作乳腺癌的递送试剂或靶向治疗剂。这种基因递送策略也是研究乳腺正常发育的一种有前途的方法。
Non-viral gene delivery vectors were developed for efficient gene transfer to hard to transfect mouse mammary epithelial cells. Ten modified versions of the same base poly(beta-amino ester), poly(1,4-butanediol diacrylate-co-5-amino-1-pentanol), were tested in both traditional 2-D monolayer and in 3-D organotypic cultures. The polymers self-assembled with plasmid DNA encoding enhanced green fluorescent protein to form nanoparticles (~100 nm) used to transfect the cells. Nanoparticle transfection efficacy was tuned by changes in synthesis and fabrication conditions and the transfection efficacy was analyzed using confocal microscopy and flow cytometry. The best performing polymeric nanoparticles transfected 57±6% of the cells in 2-D culture and 6±1% of the cells in 3-D culture. Small modifications to the polymer end-capping molecules and tuning of polymer molecular weight could either significantly enhance the transfection efficacy up to 6-fold or instead abolish efficacy completely. The efficacy of leading polymers was higher than that of the commercial transfection agent FuGENE® HD by a factor of 13 in 2-D and 2 in 3-D. These non-viral nanoparticles may be useful as delivery reagents or targeted therapeutics for breast cancer. This gene delivery strategy is also a promising approach for studying the normal development of the mammary gland.
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DOI：
10.1083/jcb.132.6.1115
发表时间：
1996-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Fialka I;Schwarz H;Reichmann E;Oft M;Busslinger M;Beug H
通讯作者：
Beug H
影响因子：
4.5
作者：
Gheisari, Y.;Soleimani, M.;Zeinali, S.
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影响因子：
29.4
作者：
Green, Jordan J.;Zugates, Gregory T.;Anderson, Daniel G.
通讯作者：
Anderson, Daniel G.
影响因子：
10.8
作者：
Green, Jordan J.;Chiu, Eugene;Anderson, Daniel G.
通讯作者：
Anderson, Daniel G.
影响因子：
2.7
作者：
Fata, Jimmie E.;Mori, Hidetoshi;Bissell, Mina J.
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Bissell, Mina J.