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Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors

OBJECTIVE(S): Human Wharton’s Jelly mesenchymal stem cells (hWMSCs) are undifferentiated cells commonly used in regenerative medicine. The aim of this study was to develop a reliable tool for tracking hWMSCs when utilized as therapeutics in burnt disorders and also to optimize the cell-based treatme...

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Autores principales: Pourfath, Mohammad Reza, Behzad-Behbahani, Abbas, Hashemi, Seyedeh Sara, Derakhsahnfar, Amin, Taheri, Mohammad Naser, Salehi, Saeede
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mashhad University of Medical Sciences 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5776439/
https://www.ncbi.nlm.nih.gov/pubmed/29372039
http://dx.doi.org/10.22038/IJBMS.2017.19783.5212
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author Pourfath, Mohammad Reza
Behzad-Behbahani, Abbas
Hashemi, Seyedeh Sara
Derakhsahnfar, Amin
Taheri, Mohammad Naser
Salehi, Saeede
author_facet Pourfath, Mohammad Reza
Behzad-Behbahani, Abbas
Hashemi, Seyedeh Sara
Derakhsahnfar, Amin
Taheri, Mohammad Naser
Salehi, Saeede
author_sort Pourfath, Mohammad Reza
collection PubMed
description OBJECTIVE(S): Human Wharton’s Jelly mesenchymal stem cells (hWMSCs) are undifferentiated cells commonly used in regenerative medicine. The aim of this study was to develop a reliable tool for tracking hWMSCs when utilized as therapeutics in burnt disorders and also to optimize the cell-based treatment procedure. MATERIALS AND METHODS: The hWMSCs were first isolated from fresh umbilical cord Wharton’s jelly and cultured. The 293LTV cell line was transfected by cGFP containing lentiviral vector and the helper plasmids for production of the viral particle. The viral particles were collected to transduce the hWMSCs. The transduced cells were finally selected based on resistance to puromycin. The burned rats (n=24) were treated with cGFP expressing hWMSCs using the cell spray method, with the cells being tracked 7, 14 and 21 days later. The rats were sacrificed 7, 14 and 21 days following treatment and paraffin embedded sections prepared from the burned area for downstream pathological analyses. RESULTS: The lentiviral particles carrying the cGFP gene were generated and the hWMSCs were transduced. The cGFP-expressing hWMSCs were detected in the burned tissue and the burned injuries were improved dramatically as compared to control. CONCLUSION: Because of the establishment of stably transduced cGFP expressing cells and the ability to detect cGFP for a relatively long-time interval, the method was found to be quite efficient for the purpose of cell tracking. The combination of hWMSC-based cell therapy and sterile Gauze Vaseline (GV) as covering was proven much more efficient than the traditional methods based on GV alone.
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spelling pubmed-57764392018-01-25 Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors Pourfath, Mohammad Reza Behzad-Behbahani, Abbas Hashemi, Seyedeh Sara Derakhsahnfar, Amin Taheri, Mohammad Naser Salehi, Saeede Iran J Basic Med Sci Original Article OBJECTIVE(S): Human Wharton’s Jelly mesenchymal stem cells (hWMSCs) are undifferentiated cells commonly used in regenerative medicine. The aim of this study was to develop a reliable tool for tracking hWMSCs when utilized as therapeutics in burnt disorders and also to optimize the cell-based treatment procedure. MATERIALS AND METHODS: The hWMSCs were first isolated from fresh umbilical cord Wharton’s jelly and cultured. The 293LTV cell line was transfected by cGFP containing lentiviral vector and the helper plasmids for production of the viral particle. The viral particles were collected to transduce the hWMSCs. The transduced cells were finally selected based on resistance to puromycin. The burned rats (n=24) were treated with cGFP expressing hWMSCs using the cell spray method, with the cells being tracked 7, 14 and 21 days later. The rats were sacrificed 7, 14 and 21 days following treatment and paraffin embedded sections prepared from the burned area for downstream pathological analyses. RESULTS: The lentiviral particles carrying the cGFP gene were generated and the hWMSCs were transduced. The cGFP-expressing hWMSCs were detected in the burned tissue and the burned injuries were improved dramatically as compared to control. CONCLUSION: Because of the establishment of stably transduced cGFP expressing cells and the ability to detect cGFP for a relatively long-time interval, the method was found to be quite efficient for the purpose of cell tracking. The combination of hWMSC-based cell therapy and sterile Gauze Vaseline (GV) as covering was proven much more efficient than the traditional methods based on GV alone. Mashhad University of Medical Sciences 2018-01 /pmc/articles/PMC5776439/ /pubmed/29372039 http://dx.doi.org/10.22038/IJBMS.2017.19783.5212 Text en Copyright: © Iranian Journal of Basic Medical Sciences http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Pourfath, Mohammad Reza
Behzad-Behbahani, Abbas
Hashemi, Seyedeh Sara
Derakhsahnfar, Amin
Taheri, Mohammad Naser
Salehi, Saeede
Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors
title Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors
title_full Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors
title_fullStr Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors
title_full_unstemmed Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors
title_short Monitoring wound healing of burn in rat model using human Wharton’s jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors
title_sort monitoring wound healing of burn in rat model using human wharton’s jelly mesenchymal stem cells containing cgfp integrated by lentiviral vectors
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5776439/
https://www.ncbi.nlm.nih.gov/pubmed/29372039
http://dx.doi.org/10.22038/IJBMS.2017.19783.5212
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