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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Autores principales:	Spoorthi Ravi Banavar, Swati Yeshwant Rawal, Ian Charles Paterson, Gurbind Singh, Fabian Davamani, Suan Phaik Khoo, Eng Lai Tan
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Mesenchymal stem cells Progenitor cells Stem cells Periodontal ligament Periodontal ligament stem cells Adult stem cells Medicine R Dentistry RK1-715
Acceso en línea:	https://doaj.org/article/f5e2a6ba125147a79606319a4bec2b17
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spelling	oai:doaj.org-article:f5e2a6ba125147a79606319a4bec2b172021-11-10T04:18:22ZEstablishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells1013-905210.1016/j.sdentj.2020.04.007https://doaj.org/article/f5e2a6ba125147a79606319a4bec2b172021-11-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S1013905219312234https://doaj.org/toc/1013-9052Mesenchymal 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, chondrogenic and adipogenic medium and then stained by Alizarin-red, Alcian-blue and Oil-Red-O respectively. MSCs from the bone-marrow were processed similarly to serve as controls. Results: The cells isolated from extracted teeth expanded successfully. On flow-cytometric analysis, 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.Spoorthi Ravi BanavarSwati Yeshwant RawalIan Charles PatersonGurbind SinghFabian DavamaniSuan Phaik KhooEng Lai TanElsevierarticleMesenchymal stem cellsProgenitor cellsStem cellsPeriodontal ligamentPeriodontal ligament stem cellsAdult stem cellsMedicineRDentistryRK1-715ENSaudi Dental Journal, Vol 33, Iss 7, Pp 693-701 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Mesenchymal stem cells Progenitor cells Stem cells Periodontal ligament Periodontal ligament stem cells Adult stem cells Medicine R Dentistry RK1-715
spellingShingle	Mesenchymal stem cells Progenitor cells Stem cells Periodontal ligament Periodontal ligament stem cells Adult stem cells Medicine R Dentistry RK1-715 Spoorthi Ravi Banavar Swati Yeshwant Rawal Ian Charles Paterson Gurbind Singh Fabian Davamani Suan Phaik Khoo Eng Lai Tan 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, chondrogenic and adipogenic medium and then stained by Alizarin-red, Alcian-blue and Oil-Red-O respectively. MSCs from the bone-marrow were processed similarly to serve as controls. Results: The cells isolated from extracted teeth expanded successfully. On flow-cytometric analysis, 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.
format	article
author	Spoorthi Ravi Banavar Swati Yeshwant Rawal Ian Charles Paterson Gurbind Singh Fabian Davamani Suan Phaik Khoo Eng Lai Tan
author_facet	Spoorthi Ravi Banavar Swati Yeshwant Rawal Ian Charles Paterson Gurbind Singh Fabian Davamani Suan Phaik Khoo Eng Lai Tan
author_sort	Spoorthi Ravi Banavar
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	https://doaj.org/article/f5e2a6ba125147a79606319a4bec2b17
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Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cells

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