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Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study

CONTEXT: It is a known fact that periodontal tissue regeneration can be achieved by the use of periodontal ligament stem cells (PDLSCs). Current mainstay of periodontal treatment is focusing on stem cell tissue engineering as an effective therapy, making it important to isolate PDLSCs from periodont...

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Autores principales: Kukreja, Bhavna Jha, Bhat, Kishore Gajanan, Kukreja, Pankaj, Kumber, Vijay Mahadev, Balakrishnan, Rajkumar, Govila, Vivek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Wolters Kluwer - Medknow 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8336774/
https://www.ncbi.nlm.nih.gov/pubmed/34393399
http://dx.doi.org/10.4103/jisp.jisp_442_20
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author Kukreja, Bhavna Jha
Bhat, Kishore Gajanan
Kukreja, Pankaj
Kumber, Vijay Mahadev
Balakrishnan, Rajkumar
Govila, Vivek
author_facet Kukreja, Bhavna Jha
Bhat, Kishore Gajanan
Kukreja, Pankaj
Kumber, Vijay Mahadev
Balakrishnan, Rajkumar
Govila, Vivek
author_sort Kukreja, Bhavna Jha
collection PubMed
description CONTEXT: It is a known fact that periodontal tissue regeneration can be achieved by the use of periodontal ligament stem cells (PDLSCs). Current mainstay of periodontal treatment is focusing on stem cell tissue engineering as an effective therapy, making it important to isolate PDLSCs from periodontal tissues. AIMS: The present research endeavor was undertaken to elucidate a technique for isolating PDLSCs for in vivo reconstructing the natural PDL tissue. SETTINGS AND DESIGN: The study design involves In vitro prospective study. MATERIALS AND METHODS: Premolar teeth were extracted from 12 patients who were under orthodontic treatment. PDL cells were scraped from their roots. Using 10 ml of Dulbecco's modified Eagle's medium with pH 7.2, the specimens of the periodontal tissue were transferred to laboratory where cell culture was done. Isolated stem cells were grown on 24-well microtiter plates-containing cover slips. They were incubated overnight at approximately 37°C in 95% air and 5% humidification. Anti-CD 45, CD73, CD90, CD105, and CD146 antibodies were used. After staining, cells were observed under phase-contrast microscopy and in inverted microscope. RESULTS: The cells showed a marked growth and 90% confluence at day 6. Cells presented thin and long fibroblastic spindle morphology. Isolated PDLSCs showed colony-forming ability at the 14(th) day after seeding. Immunohistochemical staining of PDLSCs showed positive uptake for CD146, CD90, CD73, CD105, and negative uptake for CD45. CONCLUSIONS: The human PDLSCs can be clearly isolated and characterized by using CD90, CD73, CD146, and CD105 markers of stem cells.
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spelling pubmed-83367742021-08-12 Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study Kukreja, Bhavna Jha Bhat, Kishore Gajanan Kukreja, Pankaj Kumber, Vijay Mahadev Balakrishnan, Rajkumar Govila, Vivek J Indian Soc Periodontol Original Article CONTEXT: It is a known fact that periodontal tissue regeneration can be achieved by the use of periodontal ligament stem cells (PDLSCs). Current mainstay of periodontal treatment is focusing on stem cell tissue engineering as an effective therapy, making it important to isolate PDLSCs from periodontal tissues. AIMS: The present research endeavor was undertaken to elucidate a technique for isolating PDLSCs for in vivo reconstructing the natural PDL tissue. SETTINGS AND DESIGN: The study design involves In vitro prospective study. MATERIALS AND METHODS: Premolar teeth were extracted from 12 patients who were under orthodontic treatment. PDL cells were scraped from their roots. Using 10 ml of Dulbecco's modified Eagle's medium with pH 7.2, the specimens of the periodontal tissue were transferred to laboratory where cell culture was done. Isolated stem cells were grown on 24-well microtiter plates-containing cover slips. They were incubated overnight at approximately 37°C in 95% air and 5% humidification. Anti-CD 45, CD73, CD90, CD105, and CD146 antibodies were used. After staining, cells were observed under phase-contrast microscopy and in inverted microscope. RESULTS: The cells showed a marked growth and 90% confluence at day 6. Cells presented thin and long fibroblastic spindle morphology. Isolated PDLSCs showed colony-forming ability at the 14(th) day after seeding. Immunohistochemical staining of PDLSCs showed positive uptake for CD146, CD90, CD73, CD105, and negative uptake for CD45. CONCLUSIONS: The human PDLSCs can be clearly isolated and characterized by using CD90, CD73, CD146, and CD105 markers of stem cells. Wolters Kluwer - Medknow 2021 2021-07-01 /pmc/articles/PMC8336774/ /pubmed/34393399 http://dx.doi.org/10.4103/jisp.jisp_442_20 Text en Copyright: © 2021 Indian Society of Periodontology https://creativecommons.org/licenses/by-nc-sa/4.0/This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.
spellingShingle Original Article
Kukreja, Bhavna Jha
Bhat, Kishore Gajanan
Kukreja, Pankaj
Kumber, Vijay Mahadev
Balakrishnan, Rajkumar
Govila, Vivek
Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study
title Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study
title_full Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study
title_fullStr Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study
title_full_unstemmed Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study
title_short Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study
title_sort isolation and immunohistochemical characterization of periodontal ligament stem cells: a preliminary study
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8336774/
https://www.ncbi.nlm.nih.gov/pubmed/34393399
http://dx.doi.org/10.4103/jisp.jisp_442_20
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