Recombinant sendai virus-mediated gene transfer to adipose tissue-derived stem cells (ASCs).
Adipose tissue-derived stem cells (ASCs) are expected to have clinical applications as well as other stem cells, because ASCs can be obtained safely from adult donors and used in autologous therapies without concern about rejection and the need for immunosuppression. However, the use of gene transfer with Sendai virus (SeV) vectors, which can efficiently introduce foreign genes without toxicity into several cells, with ASCs has not yet been investigated. This study documents on the use of SeV vectors for gene transfer to ASCs. The dose-dependent GFP expression of ASCs transfected with SeV vectors after 48 h of culture at 37 degrees C was first evaluated. Next, the cellular toxicity of ASCs transfected with SeV vectors was verified. In addition, SeV vectors were compared with adenovirus (AdV) vectors. Finally, the time-dependent GFP expression of ASCs transfected with SeV vectors was evaluated. The results showed that transfection of ASCs with SeV vectors results in more efficient expression of transgene (GFP expression) in the ASCs than with AdV vectors after 48 h of culture at 37 degrees C. Moreover, while the transfection of ASCs with AdV vectors at high MOIs was cytotoxic (a lot of transfected cells died) that of ASCs with SeV vectors at high MOIs was not necessarily cytotoxic. In addition, the preservation of multilineage ASCs transfected with SeV was observed. In conclusion, this is the first report describing the successful use of SeV-mediated gene transfer in ASCs, and the results indicate that SeV may thus provide advantages with respect to safety issues in gene therapy.
Yukawa
Hiroshi
H
Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Nagoya 461-0047, Japan. hiroshiy@med.nagoya-u.ac.jp
Noguchi
Hirofumi
H
Oishi
Koichi
K
Miyazaki
Takamichi
T
Kitagawa
Yasuo
Y
Inoue
Makoto
M
Hasegawa
Mamoru
M
Hayashi
Shuji
S
D000273:Adipose Tissue / Q000166:cytology* / Q000502:physiology; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology / Q000502:physiology; D000818:Animals; D002478:Cells, Cultured; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D011995:Recombination, Genetic; D029082:Sendai virus; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection; D019076:Transgenes
D049452:Green Fluorescent Proteins