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.
Lee
Chang Hyun
CH
Mirae Biotech./Cheju National University Stem Cell Research Center, Seoul 143-854, Republic of Korea.
Kim
Eun Young
EY
Jeon
Kilsoo
K
Tae
Jin Cheol
JC
Lee
Keum Sil
KS
Kim
Yeon Ok
YO
Jeong
Mi-Young
MY
Yun
Cheol-Won
CW
Jeong
Dong Kee
DK
Cho
Somi K
SK
Kim
Jae Hoon
JH
Lee
Hyo Yeon
HY
Riu
Key Zung
KZ
Cho
Ssang Goo
SG
Park
Se Pill
SP
D000818:Animals; D015415:Biomarkers; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000378:metabolism*; D017930:Genes, Reporter; D008280:Magnetics; D051379:Mice; D053758:Nanoparticles; D011094:Polyethyleneimine; D015203:Reproducibility of Results; D014162:Transfection / Q000379:methods* / Q000592:standards
D015415:Biomarkers; D011094:Polyethyleneimine