title: Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR.
abstract: 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.
authors.affiliation: Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, France; Paris-Descartes, Pierre and Marie Curie, and Paris-Diderot Universities, UFR Odontology, 75006 Paris, France; AP-HP, Hospital Complex Henri-Mondor Albert-Chenevier, CIC-BT-504, 94000 Creteil, France.
authors.forename: Ihsène
authors.initials: I
authors.lastname: Taïhi
authors.affiliation: Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, France; Paris-Descartes, Pierre and Marie Curie, and Paris-Diderot Universities, UFR Odontology, 75006 Paris, France; AP-HP, Hospital Complex Henri-Mondor Albert-Chenevier, CIC-BT-504, 94000 Creteil, France.
authors.forename: Ali
authors.initials: A
authors.lastname: Nassif
authors.affiliation: Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, France; Paris-Descartes, Pierre and Marie Curie, and Paris-Diderot Universities, UFR Odontology, 75006 Paris, France.
authors.forename: Tsouria
authors.initials: T
authors.lastname: Berbar
authors.affiliation: Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, France; Paris-Descartes, Pierre and Marie Curie, and Paris-Diderot Universities, UFR Odontology, 75006 Paris, France.
authors.forename: Juliane
authors.initials: J
authors.lastname: Isaac
authors.affiliation: Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, France; Paris-Descartes, Pierre and Marie Curie, and Paris-Diderot Universities, UFR Odontology, 75006 Paris, France; Reference Center for Dental Rare Disease, Rothschild Hospital, 75012 Paris, France.
authors.forename: Ariane
authors.initials: A
authors.lastname: Berdal
authors.affiliation: Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, France; Paris-Descartes, Pierre and Marie Curie, and Paris-Diderot Universities, UFR Odontology, 75006 Paris, France; AP-HP, Hospital Complex Henri-Mondor Albert-Chenevier, CIC-BT-504, 94000 Creteil, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Gogly
authors.affiliation: Laboratory of Molecular Oral Physiopathology, INSERM UMRS 1138, Cordeliers Research Center, 75006 Paris, France; Paris-Descartes, Pierre and Marie Curie, and Paris-Diderot Universities, UFR Odontology, 75006 Paris, France; Reference Center for Dental Rare Disease, Rothschild Hospital, 75012 Paris, France.
authors.forename: Benjamin Philippe
authors.initials: BP
authors.lastname: Fournier