Neurogenic induction of human dental pulp derived stem cells by hanging drop technique, basic fibroblast growth factor, and SHH factors.
Basic fibroblast growth factor
SHH protein
mesenchymal stem cells
neurogenesis
Journal
Dental research journal
ISSN: 1735-3327
Titre abrégé: Dent Res J (Isfahan)
Pays: Iran
ID NLM: 101471186
Informations de publication
Date de publication:
2021
2021
Historique:
received:
18
05
2020
revised:
07
08
2020
accepted:
23
01
2021
entrez:
9
9
2021
pubmed:
10
9
2021
medline:
10
9
2021
Statut:
epublish
Résumé
The progressive destruction of nerve cells in nervous system will induce neurodegenerative diseases. Recently, cell-based therapies have attracted the attention of researchers in the treatment of these abnormal conditions. Thus, the aim of this study was to provide a simple and efficient way to differentiate human dental pulp stem cells into neural cell-like to achieve a homogeneous population of these cells for transplantation in neurodegenerative diseases. In this basic research, human dental pulp stem cells were isolated and characterized by immunocytochemistry and flow cytometry techniques. In the following, the cells were cultured using hanging drop as three-dimensional (3D) and tissue culture plate as 2D techniques. Subsequently, cultured cells were differentiated into neuron cell-like in the presence of FGF and Sonic hedgehog (SHH) factors. Finally, the percentage of cells expressing Neu N and β tubulin III markers was determined using immunocytochemistry technique. Finally, all data were analyzed using the SPSS software. Flow cytometry and immunocytochemistry results indicated that human dental pulp-derived stem cells were CD90, CD106-positive, but were negative for CD34, CD45 markers ( It is concluded that due to the wide range of SHH functions and the facilitation of intercellular connections in the hanging droop method, it is recommended that the use of hanging drop method and SHH factor can be effective in increasing the efficiency of cell differentiation.
Sections du résumé
BACKGROUND
BACKGROUND
The progressive destruction of nerve cells in nervous system will induce neurodegenerative diseases. Recently, cell-based therapies have attracted the attention of researchers in the treatment of these abnormal conditions. Thus, the aim of this study was to provide a simple and efficient way to differentiate human dental pulp stem cells into neural cell-like to achieve a homogeneous population of these cells for transplantation in neurodegenerative diseases.
MATERIALS AND METHODS
METHODS
In this basic research, human dental pulp stem cells were isolated and characterized by immunocytochemistry and flow cytometry techniques. In the following, the cells were cultured using hanging drop as three-dimensional (3D) and tissue culture plate as 2D techniques. Subsequently, cultured cells were differentiated into neuron cell-like in the presence of FGF and Sonic hedgehog (SHH) factors. Finally, the percentage of cells expressing Neu N and β tubulin III markers was determined using immunocytochemistry technique. Finally, all data were analyzed using the SPSS software.
RESULTS
RESULTS
Flow cytometry and immunocytochemistry results indicated that human dental pulp-derived stem cells were CD90, CD106-positive, but were negative for CD34, CD45 markers (
CONCLUSION
CONCLUSIONS
It is concluded that due to the wide range of SHH functions and the facilitation of intercellular connections in the hanging droop method, it is recommended that the use of hanging drop method and SHH factor can be effective in increasing the efficiency of cell differentiation.
Types de publication
Journal Article
Langues
eng
Pagination
57Informations de copyright
Copyright: © 2021 Dental Research Journal.
Déclaration de conflit d'intérêts
The authors of this manuscript declare that they have no conflicts of interest, real or perceived, financial or nonfinancial in this article.
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