Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR
Gingival stem cells (GSCs) are recently isolated multipotent cells. Their osteogenic capacity has been validated in vitro and may be transferred to human cell therapy for maxillary large bone defects, as they share a neural crest cell origin with jaw bone cells. RT-qPCR is a widely used technique to...
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| Format: | Article |
| Language: | English |
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Wiley
2016-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2016/6261490 |
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| author | Ihsène Taïhi Ali Nassif Tsouria Berbar Juliane Isaac Ariane Berdal Bruno Gogly Benjamin Philippe Fournier |
| author_facet | Ihsène Taïhi Ali Nassif Tsouria Berbar Juliane Isaac Ariane Berdal Bruno Gogly Benjamin Philippe Fournier |
| author_sort | Ihsène Taïhi |
| collection | DOAJ |
| description | Gingival stem cells (GSCs) are recently isolated multipotent cells. Their osteogenic capacity has been validated in vitro and may be transferred to human cell therapy for maxillary large bone defects, as they share a neural crest cell origin with jaw bone cells. RT-qPCR is a widely used technique to study gene expression and may help us to follow osteoblast differentiation of GSCs. For accurate results, the choice of reliable housekeeping genes (HKGs) is crucial. The aim of this study was to select the most reliable HKGs for GSCs study and their osteogenic differentiation (dGSCs). The analysis was performed with ten selected HKGs using four algorithms: ΔCt comparative method, GeNorm, BestKeeper, and NormFinder. This study demonstrated that three HKGs, SDHA, ACTB, and B2M, were the most stable to study GSC, whereas TBP, SDHA, and ALAS1 were the most reliable to study dGSCs. The comparison to stem cells of mesenchymal origin (ASCs) showed that SDHA/HPRT1 were the most appropriate for ASCs study. The choice of suitable HKGs for GSCs is important as it gave access to an accurate analysis of osteogenic differentiation. It will allow further study of this interesting stem cells source for future human therapy. |
| format | Article |
| id | doaj-art-c1dcacd154564463bd698345b038666e |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-c1dcacd154564463bd698345b038666e2025-02-03T01:25:30ZengWileyStem Cells International1687-966X1687-96782016-01-01201610.1155/2016/62614906261490Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCRIhsène Taïhi0Ali Nassif1Tsouria Berbar2Juliane Isaac3Ariane Berdal4Bruno Gogly5Benjamin Philippe Fournier6Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, FranceLaboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, FranceLaboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, FranceLaboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, FranceLaboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, FranceLaboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, FranceLaboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, FranceGingival stem cells (GSCs) are recently isolated multipotent cells. Their osteogenic capacity has been validated in vitro and may be transferred to human cell therapy for maxillary large bone defects, as they share a neural crest cell origin with jaw bone cells. RT-qPCR is a widely used technique to study gene expression and may help us to follow osteoblast differentiation of GSCs. For accurate results, the choice of reliable housekeeping genes (HKGs) is crucial. The aim of this study was to select the most reliable HKGs for GSCs study and their osteogenic differentiation (dGSCs). The analysis was performed with ten selected HKGs using four algorithms: ΔCt comparative method, GeNorm, BestKeeper, and NormFinder. This study demonstrated that three HKGs, SDHA, ACTB, and B2M, were the most stable to study GSC, whereas TBP, SDHA, and ALAS1 were the most reliable to study dGSCs. The comparison to stem cells of mesenchymal origin (ASCs) showed that SDHA/HPRT1 were the most appropriate for ASCs study. The choice of suitable HKGs for GSCs is important as it gave access to an accurate analysis of osteogenic differentiation. It will allow further study of this interesting stem cells source for future human therapy.http://dx.doi.org/10.1155/2016/6261490 |
| spellingShingle | Ihsène Taïhi Ali Nassif Tsouria Berbar Juliane Isaac Ariane Berdal Bruno Gogly Benjamin Philippe Fournier Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR Stem Cells International |
| title | Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR |
| title_full | Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR |
| title_fullStr | Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR |
| title_full_unstemmed | Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR |
| title_short | Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR |
| title_sort | validation of housekeeping genes to study human gingival stem cells and their in vitro osteogenic differentiation using real time rt qpcr |
| url | http://dx.doi.org/10.1155/2016/6261490 |
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