Green Jordan J, Zhou Betty Y, Mitalipova Maisam M, Beard Caroline, Langer Robert, Jaenisch Rudolf, Anderson Daniel G
Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
Nano Lett. 2008 Oct;8(10):3126-30. doi: 10.1021/nl8012665. Epub 2008 Aug 29.
We develop biodegradable polymeric nanoparticles to facilitate nonviral gene transfer to human embryonic stem cells (hESCs). Small (approximately 200 nm), positively charged (approximately 10 mV) particles are formed by the self assembly of cationic, hydrolytically degradable poly(beta-amino esters) and plasmid DNA. By varying the end group of the polymer, we can tune the biophysical properties of the resulting nanoparticles and their gene-delivery efficacy. We created an OCT4-driven GFP hES cell line to allow the rapid identification of nanoparticles that facilitate gene transfer while maintaining an hESC undifferentiated state. Using this cell system, we synthesized nanoparticles that have gene delivery efficacy that is up to 4 times higher than that of the leading commercially available transfection agent, Lipofectamine 2000. Importantly, these materials have minimal toxicity and do not adversely affect hESC colony morphology or cause nonspecific differentiation.
我们开发了可生物降解的聚合物纳米颗粒，以促进非病毒基因向人类胚胎干细胞（hESCs）的转移。阳离子型、可水解降解的聚（β-氨基酯）和质粒DNA通过自组装形成了小的（约200纳米）、带正电的（约10毫伏）颗粒。通过改变聚合物的端基，我们可以调节所得纳米颗粒的生物物理性质及其基因递送效率。我们创建了一个由OCT4驱动的绿色荧光蛋白hES细胞系，以便快速识别促进基因转移同时维持hESC未分化状态的纳米颗粒。利用这个细胞系统，我们合成了基因递送效率比领先的市售转染试剂Lipofectamine 2000高多达4倍的纳米颗粒。重要的是，这些材料毒性极小，不会对hESC集落形态产生不利影响，也不会导致非特异性分化。
