Reference - Detail
| RRC ID | 36769 |
|---|---|
| Author | Park E, Cho HB, Takimoto K. |
| Title | Effective gene delivery into adipose-derived stem cells: transfection of cells in suspension with the use of a nuclear localization signal peptide-conjugated polyethylenimine. |
| Journal | Cytotherapy |
| Abstract |
BACKGROUND AIMS:Adipose-derived stem cells have the ability to turn into several clinically important cell types. However, it is difficult to transfect these cells with the use of conventional cationic lipid-based reagents. Polyethylenimine (PEI) is considered to be an inexpensive and effective tool for delivery of nucleic acids into mammalian cells. METHODS:We used a linear PEI conjugated with the nuclear localization signal (NLS) peptide of Simian vacuolating virus 40 large T antigen (PEI-NLS) for transfection of plasmid DNA into adipose-derived cells. We also tested if transfection of cells in suspension might improve the degree and duration of exogenous gene expression. RESULTS:Transfection of cells in suspension with the use of a PEI conjugated with an NLS peptide resulted in high levels of reporter gene expression for an extended period of time in clonal 3T3-L1 preadipocytes and native human adipose-derived stem cells. The reporter gene expression increased for 3 days after the addition of the PEI-NLS peptide-DNA mixture in cell suspension and remained significant for at least 7 days. Cell density did not influence the level of reporter gene expression. Thus, the suspension method with the use of an NLS peptide-conjugated PEI leads to a robust and sustained expression of exogenous genes in adipose-derived cells. CONCLUSIONS:The devised transfection method may be useful for reprogramming of adipose-derived stem cells and cell-based therapy. |
| Volume | 17(5) |
| Pages | 536-42 |
| Published | 2015-5-1 |
| DOI | 10.1016/j.jcyt.2014.11.008 |
| PII | S1465-3249(14)00879-2 |
| PMID | 25618562 |
| MeSH | 3T3-L1 Cells Adipose Tissue / cytology* Amino Acid Sequence Animals CHO Cells Cell Count Cricetinae Cricetulus Gene Expression Genes, Reporter HeLa Cells Humans Mice Molecular Sequence Data Nuclear Localization Signals / chemistry Nuclear Localization Signals / metabolism* Peptides / chemistry Peptides / metabolism* Polyethyleneimine / chemistry* Stem Cells / metabolism* Transfection / methods* |
| IF | 4.218 |
| Times Cited | 19 |
| WOS Category | MEDICINE, RESEARCH & EXPERIMENTAL BIOTECHNOLOGY & APPLIED MICROBIOLOGY CELL & TISSUE ENGINEERING HEMATOLOGY CELL BIOLOGY |
| Resource | |
| Human and Animal Cells | C2C12(RCB0987) |