title: Nanoparticles for gene transfer to human embryonic stem cell colonies.
abstract: 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.
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Jordan J
authors.initials: JJ
authors.lastname: Green
authors.forename: Betty Y
authors.initials: BY
authors.lastname: Zhou
authors.forename: Maisam M
authors.initials: MM
authors.lastname: Mitalipova
authors.forename: Caroline
authors.initials: C
authors.lastname: Beard
authors.forename: Robert
authors.initials: R
authors.lastname: Langer
authors.forename: Rudolf
authors.initials: R
authors.lastname: Jaenisch
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Anderson