A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell

Mesenchymal stem cells (MSCs) hold tremendous potential for therapeutic use in stem cell-based gene therapy. Ex vivo genetic modification of MSCs with beneficial genes of interest is a prerequisite for successful use of stem cell-based therapeutic applications. However, genetic manipulation of MS...

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Main Authors: Abdul Halim, Nur Shuhaidatul Sarmiza, Shaik Fakiruddin, Kamal, Ali, Syed Atif, Yahaya, Badrul Hisham
Format: Article
Language:English
Published: MDPI 2014
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Online Access:http://eprints.usm.my/37995/1/A_Comparative_Study_of_Non-Viral_Gene_Delivery_Techniques.pdf
http://eprints.usm.my/37995/
https://doi.org/10.3390/ijms150915044
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spelling my.usm.eprints.37995 http://eprints.usm.my/37995/ A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell Abdul Halim, Nur Shuhaidatul Sarmiza Shaik Fakiruddin, Kamal Ali, Syed Atif Yahaya, Badrul Hisham RK1-715 Dentistry Mesenchymal stem cells (MSCs) hold tremendous potential for therapeutic use in stem cell-based gene therapy. Ex vivo genetic modification of MSCs with beneficial genes of interest is a prerequisite for successful use of stem cell-based therapeutic applications. However, genetic manipulation of MSCs is challenging because they are resistant to commonly used methods to introduce exogenous DNA or RNA. Herein we compared the effectiveness of several techniques (classic calcium phosphate precipitation, cationic polymer, and standard electroporation) with that of microporation technology to introduce the plasmid encoding for angiopoietin-1 (ANGPT-1) and enhanced green fluorescent protein (eGFP) into human adipose-derived MSCs (hAD-MSCs). The microporation technique had a higher transfection efficiency, with up to 50% of the viable hAD-MSCs being transfected, compared to the other transfection techniques, for which less than 1% of cells were positive for eGFP expression following transfection. The capability of cells to proliferate and differentiate into three major lineages (chondrocytes, adipocytes, and osteocytes) was found to be independent of the technique used for transfection. These results show that the microporation technique is superior to the others in terms of its ability to transfect hAD-MSCs without affecting their proliferation and differentiation capabilities. Therefore, this study provides a foundation for the selection of techniques when using ex vivo gene manipulation for cell-based gene therapy with MSCs as the vehicle for gene delivery. MDPI 2014 Article PeerReviewed application/pdf en http://eprints.usm.my/37995/1/A_Comparative_Study_of_Non-Viral_Gene_Delivery_Techniques.pdf Abdul Halim, Nur Shuhaidatul Sarmiza and Shaik Fakiruddin, Kamal and Ali, Syed Atif and Yahaya, Badrul Hisham (2014) A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell. International Journal of Molecular Sciences, 15 (9). pp. 15044-15060. ISSN 1422-0067 https://doi.org/10.3390/ijms150915044
institution Universiti Sains Malaysia
building Hamzah Sendut Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Sains Malaysia
content_source USM Institutional Repository
url_provider http://eprints.usm.my/
language English
topic RK1-715 Dentistry
spellingShingle RK1-715 Dentistry
Abdul Halim, Nur Shuhaidatul Sarmiza
Shaik Fakiruddin, Kamal
Ali, Syed Atif
Yahaya, Badrul Hisham
A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell
description Mesenchymal stem cells (MSCs) hold tremendous potential for therapeutic use in stem cell-based gene therapy. Ex vivo genetic modification of MSCs with beneficial genes of interest is a prerequisite for successful use of stem cell-based therapeutic applications. However, genetic manipulation of MSCs is challenging because they are resistant to commonly used methods to introduce exogenous DNA or RNA. Herein we compared the effectiveness of several techniques (classic calcium phosphate precipitation, cationic polymer, and standard electroporation) with that of microporation technology to introduce the plasmid encoding for angiopoietin-1 (ANGPT-1) and enhanced green fluorescent protein (eGFP) into human adipose-derived MSCs (hAD-MSCs). The microporation technique had a higher transfection efficiency, with up to 50% of the viable hAD-MSCs being transfected, compared to the other transfection techniques, for which less than 1% of cells were positive for eGFP expression following transfection. The capability of cells to proliferate and differentiate into three major lineages (chondrocytes, adipocytes, and osteocytes) was found to be independent of the technique used for transfection. These results show that the microporation technique is superior to the others in terms of its ability to transfect hAD-MSCs without affecting their proliferation and differentiation capabilities. Therefore, this study provides a foundation for the selection of techniques when using ex vivo gene manipulation for cell-based gene therapy with MSCs as the vehicle for gene delivery.
format Article
author Abdul Halim, Nur Shuhaidatul Sarmiza
Shaik Fakiruddin, Kamal
Ali, Syed Atif
Yahaya, Badrul Hisham
author_facet Abdul Halim, Nur Shuhaidatul Sarmiza
Shaik Fakiruddin, Kamal
Ali, Syed Atif
Yahaya, Badrul Hisham
author_sort Abdul Halim, Nur Shuhaidatul Sarmiza
title A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell
title_short A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell
title_full A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell
title_fullStr A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell
title_full_unstemmed A comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell
title_sort comparative study of non-viral gene delivery techniques to human adipose-derived mesenchymal stem cell
publisher MDPI
publishDate 2014
url http://eprints.usm.my/37995/1/A_Comparative_Study_of_Non-Viral_Gene_Delivery_Techniques.pdf
http://eprints.usm.my/37995/
https://doi.org/10.3390/ijms150915044
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score 13.209306