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 transfe...

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Main Authors: Hiroshi Yukawa, Hirofumi Noguchi, Koichi Oishi, Takamichi Miyazaki, Yasuo Kitagawa, Makoto Inoue, Mamoru Hasegawa, Shuji Hayashi
Format: Article
Language:English
Published: SAGE Publishing 2008-01-01
Series:Cell Transplantation
Online Access:https://doi.org/10.3727/000000008783907071
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author Hiroshi Yukawa
Hirofumi Noguchi
Koichi Oishi
Takamichi Miyazaki
Yasuo Kitagawa
Makoto Inoue
Mamoru Hasegawa
Shuji Hayashi
author_facet Hiroshi Yukawa
Hirofumi Noguchi
Koichi Oishi
Takamichi Miyazaki
Yasuo Kitagawa
Makoto Inoue
Mamoru Hasegawa
Shuji Hayashi
author_sort Hiroshi Yukawa
collection DOAJ
description 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°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°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.
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spelling doaj.art-4f50f9384dbf4069a94b3ba77b13f09a2022-12-21T19:55:41ZengSAGE PublishingCell Transplantation0963-68971555-38922008-01-011710.3727/000000008783907071Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)Hiroshi Yukawa0Hirofumi Noguchi1Koichi Oishi2Takamichi Miyazaki3Yasuo Kitagawa4Makoto Inoue5Mamoru Hasegawa6Shuji Hayashi7Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Nagoya 461-0047, JapanDepartment of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Nagoya 461-0047, JapanDepartment of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Nagoya 461-0047, JapanDepartment of Bioagricultural Sciences, Nagoya University Graduate School of Bioagricultural Sciences, Nagoya 464-8601, JapanDepartment of Bioagricultural Sciences, Nagoya University Graduate School of Bioagricultural Sciences, Nagoya 464-8601, JapanDNAVEC Corporation, Ibaraki 305-0856, JapanDNAVEC Corporation, Ibaraki 305-0856, JapanDepartment of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Nagoya 461-0047, JapanAdipose 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°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°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.https://doi.org/10.3727/000000008783907071
spellingShingle Hiroshi Yukawa
Hirofumi Noguchi
Koichi Oishi
Takamichi Miyazaki
Yasuo Kitagawa
Makoto Inoue
Mamoru Hasegawa
Shuji Hayashi
Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)
Cell Transplantation
title Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)
title_full Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)
title_fullStr Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)
title_full_unstemmed Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)
title_short Recombinant Sendai Virus-Mediated Gene Transfer to Adipose Tissue-Derived Stem Cells (ASCs)
title_sort recombinant sendai virus mediated gene transfer to adipose tissue derived stem cells ascs
url https://doi.org/10.3727/000000008783907071
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