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Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells

Mesenchymal stem cells (MSCs) are extensively used in tissue regenerative procedures. One source of MSCs is the periodontal ligament (PDL) of teeth. Isolation of MSCs from extracted teeth is reasonably simple, being less invasive and presenting fewer ethical concerns than does the harvesting of MSC&...

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Main Authors: Banavar, Spoorthi Ravi, Rawal, Swati Yeshwant, Paterson, Ian Charles, Singh, Gurbind, Davamani, Fabian, Khoo, Suan Phaik, Tan, Eng Lai
Format: Article
Published: Elsevier 2021
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R Medicine
Online Access:http://eprints.um.edu.my/34901/
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spelling my.um.eprints.349012022-05-26T04:05:54Z http://eprints.um.edu.my/34901/ Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells Banavar, Spoorthi Ravi Rawal, Swati Yeshwant Paterson, Ian Charles Singh, Gurbind Davamani, Fabian Khoo, Suan Phaik Tan, Eng Lai R Medicine Mesenchymal stem cells (MSCs) are extensively used in tissue regenerative procedures. One source of MSCs is the periodontal ligament (PDL) of teeth. Isolation of MSCs from extracted teeth is reasonably simple, being less invasive and presenting fewer ethical concerns than does the harvesting of MSC's from other sites. The objectives of this study were to isolate and characterize the PDL stem cells (PDLSC) from healthy adults' extracted teeth and then to characterize them by comparing them with bone-marrow derived MSCs (BMMSC). Methods: The PDL tissue was scraped from the roots of freshly extracted teeth to enzymatically digest using collagenase. The cells were sub-cultured. Flow-cytometric analysis for the MSC surface-markers CD105, CD73, CD166, CD90, CD34, CD45 and HLA-DR was performed. To confirm the phenotype, total RNA was extracted to synthesize cDNA and which was then subjected to RT-PCR. The gene-expression for Oct4A, Sox2, NANOG and GAPDH was determined by gel-electrophoresis. To assess their multilineage potential, cells were cultured with osteogenic, chondro-genic and adipogenic medium and then stained by Alizarin-red, Alcian-blue and Oil-Red-O respec-tively. MSCs from the bone-marrow were processed similarly to serve as controls. Results: The cells isolated from extracted teeth expanded successfully. On flow-cytometric anal-ysis, the cells were positive for CD73, CD90, CD105, CD166 and negative for CD34, CD45 and HLA-DR. The PDLSCs expressed Oct4A, Sox2, and NANOG mRNA with GAPDH expression. Cells cultured in the osteogenic, chondrogenic and adipogenic media stained positive for Alizarin red, Alcian-blue and Oil-Red-O respectively. The surface marker expression and the trilineage differentiation characteristics were comparable to those of the BMMSCs. Conclusions: The periodontal ligament tissue of extracted teeth is a potential source of therapeutically useful MSCs. Harvesting them is not invasive and are a promising source of MSC as the PDLSCs showed characteristics similar to those of the highly regarded MSC's derived from bone-marrow. (C) 2020 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. Elsevier 2021-11 Article PeerReviewed Banavar, Spoorthi Ravi and Rawal, Swati Yeshwant and Paterson, Ian Charles and Singh, Gurbind and Davamani, Fabian and Khoo, Suan Phaik and Tan, Eng Lai (2021) Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells. Saudi Dental Journal, 33 (7). pp. 693-701. ISSN 1013-9052, DOI https://doi.org/10.1016/j.sdentj.2020.04.007 <https://doi.org/10.1016/j.sdentj.2020.04.007>. 10.1016/j.sdentj.2020.04.007
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
topic R Medicine
spellingShingle R Medicine
Banavar, Spoorthi Ravi
Rawal, Swati Yeshwant
Paterson, Ian Charles
Singh, Gurbind
Davamani, Fabian
Khoo, Suan Phaik
Tan, Eng Lai
Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells
description Mesenchymal stem cells (MSCs) are extensively used in tissue regenerative procedures. One source of MSCs is the periodontal ligament (PDL) of teeth. Isolation of MSCs from extracted teeth is reasonably simple, being less invasive and presenting fewer ethical concerns than does the harvesting of MSC's from other sites. The objectives of this study were to isolate and characterize the PDL stem cells (PDLSC) from healthy adults' extracted teeth and then to characterize them by comparing them with bone-marrow derived MSCs (BMMSC). Methods: The PDL tissue was scraped from the roots of freshly extracted teeth to enzymatically digest using collagenase. The cells were sub-cultured. Flow-cytometric analysis for the MSC surface-markers CD105, CD73, CD166, CD90, CD34, CD45 and HLA-DR was performed. To confirm the phenotype, total RNA was extracted to synthesize cDNA and which was then subjected to RT-PCR. The gene-expression for Oct4A, Sox2, NANOG and GAPDH was determined by gel-electrophoresis. To assess their multilineage potential, cells were cultured with osteogenic, chondro-genic and adipogenic medium and then stained by Alizarin-red, Alcian-blue and Oil-Red-O respec-tively. MSCs from the bone-marrow were processed similarly to serve as controls. Results: The cells isolated from extracted teeth expanded successfully. On flow-cytometric anal-ysis, the cells were positive for CD73, CD90, CD105, CD166 and negative for CD34, CD45 and HLA-DR. The PDLSCs expressed Oct4A, Sox2, and NANOG mRNA with GAPDH expression. Cells cultured in the osteogenic, chondrogenic and adipogenic media stained positive for Alizarin red, Alcian-blue and Oil-Red-O respectively. The surface marker expression and the trilineage differentiation characteristics were comparable to those of the BMMSCs. Conclusions: The periodontal ligament tissue of extracted teeth is a potential source of therapeutically useful MSCs. Harvesting them is not invasive and are a promising source of MSC as the PDLSCs showed characteristics similar to those of the highly regarded MSC's derived from bone-marrow. (C) 2020 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.
format Article
author Banavar, Spoorthi Ravi
Rawal, Swati Yeshwant
Paterson, Ian Charles
Singh, Gurbind
Davamani, Fabian
Khoo, Suan Phaik
Tan, Eng Lai
author_facet Banavar, Spoorthi Ravi
Rawal, Swati Yeshwant
Paterson, Ian Charles
Singh, Gurbind
Davamani, Fabian
Khoo, Suan Phaik
Tan, Eng Lai
author_sort Banavar, Spoorthi Ravi
title Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells
title_short Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells
title_full Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells
title_fullStr Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells
title_full_unstemmed Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells
title_sort establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells
publisher Elsevier
publishDate 2021
url http://eprints.um.edu.my/34901/
_version_ 1735409631952371712
score 13.211869

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