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’...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-11-01
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| Series: | Saudi Dental Journal |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S1013905219312234 |
| _version_ | 1818931941348474880 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-20T04:24:35Z |
| format | Article |
| id | doaj.art-f5e2a6ba125147a79606319a4bec2b17 |
| institution | Directory Open Access Journal |
| issn | 1013-9052 |
| language | English |
| last_indexed | 2024-12-20T04:24:35Z |
| publishDate | 2021-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Saudi Dental Journal |
| spelling | doaj.art-f5e2a6ba125147a79606319a4bec2b172022-12-21T19:53:33ZengElsevierSaudi Dental Journal1013-90522021-11-01337693701Establishing a technique for isolation and characterization of human periodontal ligament derived mesenchymal stem cellsSpoorthi Ravi Banavar0Swati Yeshwant Rawal1Ian Charles Paterson2Gurbind Singh3Fabian Davamani4Suan Phaik Khoo5Eng Lai Tan6Oral Diagnostic and Surgical Sciences, School of Dentistry, International Medical University, Kuala Lumpur, Malaysia; Corresponding author.Department of Surgical Sciences, Marquette University, USAFaculty of Dentistry, University of Malaya, Kuala Lumpur, MalaysiaSchool of Pharmacy, International Medical University, Kuala Lumpur, MalaysiaSchool of Health Sciences, International Medical University, Kuala Lumpur, MalaysiaOral Diagnostic and Surgical Sciences, School of Dentistry, International Medical University, Kuala Lumpur, MalaysiaSchool of Pharmacy, International Medical University, Kuala Lumpur, MalaysiaMesenchymal 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.http://www.sciencedirect.com/science/article/pii/S1013905219312234Mesenchymal stem cellsProgenitor cellsStem cellsPeriodontal ligamentPeriodontal ligament stem cellsAdult stem cells |
| spellingShingle | 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 Saudi Dental Journal Mesenchymal stem cells Progenitor cells Stem cells Periodontal ligament Periodontal ligament stem cells Adult stem cells |
| title | 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_short | 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 |
| topic | Mesenchymal stem cells Progenitor cells Stem cells Periodontal ligament Periodontal ligament stem cells Adult stem cells |
| url | http://www.sciencedirect.com/science/article/pii/S1013905219312234 |
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