In vitro comparison of the biological effects of three transfection methods for magnetically labeling mouse embryonic stem cells with ferumoxides.
In vivo MRI of stem cells (SCs) is an emerging application to evaluate the role of cell therapy in restoring the injured myocardium. The high spatial and temporal resolution combined with iron-oxide-based intracellular labeling techniques will provide a sensitive, noninvasive, dual imaging modality for both cells and myocardium. In order to facilitate this novel imaging approach, much effort has been directed towards developing efficient transfection methods. While techniques utilizing poly-L-lysine (PLL), protamine sulfate (PS), and electroporation (ELP) have been proposed, the fundamental biological effects of these methods on mouse embryonic SCs (mESC) have not been investigated systematically. In this study a longitudinal in vitro evaluation of cellular viability, apoptosis, proliferation, and cardiac differentiation of magnetically labeled mESC was conducted. No significant difference was seen in these biological parameters among the three transfection methods. However, cardiac differentiation was most attenuated by ELP, and iron uptake was most effective by PS.
Suzuki
Yoriyasu
Y
Division of Cardiovascular Medicine, Stanford University, Stanford, CA 94305-5233, USA.
Zhang
Sally
S
Kundu
Pratima
P
Yeung
Alan C
AC
Robbins
Robert C
RC
Yang
Phillip C
PC
D000464:Alginates; D000704:Analysis of Variance; D000818:Animals; D017209:Apoptosis; D003287:Contrast Media / Q000493:pharmacokinetics*; D003911:Dextrans; D018274:Electroporation; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology*; D052203:Ferrosoferric Oxide; D066298:In Vitro Techniques; D007501:Iron / Q000493:pharmacokinetics*; D008279:Magnetic Resonance Imaging; D058185:Magnetite Nanoparticles; D051379:Mice; D010087:Oxides / Q000493:pharmacokinetics*; D011107:Polylysine / Q000031:analogs & derivatives; D011479:Protamines; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*
D000464:Alginates; D003287:Contrast Media; D003911:Dextrans; D058185:Magnetite Nanoparticles; D010087:Oxides; D011479:Protamines; D017382:Reactive Oxygen Species; C051546:alginate-polylysine-alginate; D011107:Polylysine; D007501:Iron; C065507:ferumoxides; D052203:Ferrosoferric Oxide