Chang Hyun Lee

Simple, Efficient, and Reproducible Gene Transfection of Mouse Embryonic Stem Cells by Magnetofection

Embryonic stem (ES) cells are recognized as an excellent cell culture model for studying developmental mechanisms and their therapeutic modulations. The aim of this work was to define whether using magnetofection was an efficient way to manipulate stem cells genetically without adversely affecting their proliferation or self-renewal capacity. We compared our magnetofection results to those of a conservative method using FuGENE 6. Using enhanced green fluorescent protein (eGFP) as a reporter gene in D3 mouse ES (mES) cells, we found that magnetofection gave a significantly higher efficiency (45%) of gene delivery in stem cells than did the FuGENE 6 method (15%), whereas both demonstrated efficient transfection in NIH-3T3 cells (60%). Although the transfected D3 (D3-eGFP) mES cells had undergone a large number of passages (>50), a high percentage of cells retained ES markers such as Oct-4 and stage-specific embryonic antigen-1 (SSEA-1). They also retained the ability to form embryoid bodies and differentiated in vitro into cells of the three germ layers. eGFP expression was sustained during stem cell proliferation and differentiation. This is the first transfection report using magnetofection in ES cells. On the basis of our results, we conclude that magnetofection is an efficient and reliable method for the introduction of foreign DNA into mouse ES cells and may become the method of choice.

Synthesis of Au-BiVO4 nanocomposite through anodic electrodeposition followed by galvanic replacement and its application to the photocatalytic decomposition of methyl orange.

A Au-BiVO(4) nanocomposite is synthesized by a two-step strategy involving anodic electrodeposition combined with in situ galvanic replacement. First, a BiVO(4) layer is prepared by the anodic oxidation of pre-electrodeposited Bi film in a VO(4)(3-) containing electrolyte. Thus-prepared BiVO(4) film contains excess metallic Bi, which is then galvanically replaced with Au from an aqueous HAuCl(4) solution, resulting in the Au-BiVO(4) composite in the second step. Optical, photoelectrochemical and photocatalytic properties are investigated by using X-ray diffraction, energy-dispersive X-ray analysis, diffuse reflectance spectrometry, and photoelectrochemical analyses. The visible-light photocatalytic activity of the Au-BiVO(4) composite is evaluated using the decomposition of methyl orange dye and is superior to the bare BiVO(4) film counterpart.