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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| Format: | Article |
| Language: | English |
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MDPI
2014
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| Online Access: | http://eprints.usm.my/37995/1/A_Comparative_Study_of_Non-Viral_Gene_Delivery_Techniques.pdf |
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| 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 |
| collection | USM |
| 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. |
| first_indexed | 2024-03-06T15:12:43Z |
| format | Article |
| id | usm.eprints-37995 |
| institution | Universiti Sains Malaysia |
| language | English |
| last_indexed | 2024-03-06T15:12:43Z |
| publishDate | 2014 |
| publisher | MDPI |
| record_format | dspace |
| spelling | usm.eprints-379952017-12-20T00:53:17Z 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 |
| 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 |
| title | 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_short | 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 |
| topic | RK1-715 Dentistry |
| url | http://eprints.usm.my/37995/1/A_Comparative_Study_of_Non-Viral_Gene_Delivery_Techniques.pdf |
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