Microporation is a valuable transfection method for gene expression in human adipose tissue-derived stem cells.
Stem cells are a promising resource for gene therapy. Adipose tissue-derived stem cells (ADSCs) offer advantages because of their abundance and ease of isolation. However, it is difficult to transduce genes into ADSCs by common transfection methods, especially nonviral methods. We report here the use of a new electroporation method, termed "microporation," to transduce plasmids into human ADSCs (hADSCs). We determined optimal conditions that led to efficient transfection of >76.1% of the microporated hADSCs with only minimal cell damage or cytotoxicity. We demonstrated the expression of both enhanced green fluorescent protein (EGFP) and luciferase from different promoters in microporated hADSCs. More important, the microporated hADSCs retained their multipotency and reporter gene expression was maintained for >2 weeks in vitro and in vivo. We further showed that a Tet-ON-inducible gene expression system could be microporated into hADSCs and that this system was functional following transplantation of the microporated cells into nude mice. Taken together, our data demonstrate that microporation allows a highly efficient transfection of hADSCs, without impairing their stem cell properties.
Wang
Yan-Hsiung
YH
Orthopaedic Research Center, Kaohsiung Medical University, Kaohsiung, Taiwan.
Ho
Mei-Ling
ML
Chang
Je-Ken
JK
Chu
Hai-Chang
HC
Lai
Shu-Chin
SC
Wang
Gwo-Jaw
GJ
D000273:Adipose Tissue / Q000166:cytology*; D015153:Blotting, Western; D002454:Cell Differentiation / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008156:Luciferases