title: Immunophenotyping Reveals the Diversity of Human Dental Pulp Mesenchymal Stromal CellsIn vivoand Their Evolution uponIn vitroAmplification.
abstract: Mesenchymal stromal/stem cells (MSCs) from human dental pulp (DP) can be expandedin vitrofor cell-based and regenerative dentistry therapeutic purposes. However, their heterogeneity may be a hurdle to the achievement of reproducible and predictable therapeutic outcomes. To get a better knowledge about this heterogeneity, we designed a flow cytometric strategy to analyze the phenotype of DP cellsin vivoand uponin vitroexpansion with stem cell markers. We focused on the CD31-cell population to exclude endothelial and leukocytic cells. Results showed that thein vivoCD31-DP cell population contained 1.4% of CD56+, 1.5% of CD146+, 2.4% of CD271+and 6.3% of MSCA-1+cells but very few Stro-1+cells (≤ 1%). CD56+, CD146+, CD271+, and MSCA-1+cell subpopulations expressed various levels of these markers. CD146+MSCA-1+, CD271+MSCA-1+, and CD146+CD271+cells were the most abundant DP-MSC populations. Analysis of DP-MSCs expandedin vitrowith a medicinal manufacturing approach showed that CD146 was expressed by about 50% of CD56+, CD271+, MSCA-1+, and Stro-1+cells, and MSCA-1 by 15-30% of CD56+, CD146+, CD271+, and Stro-1+cells. These ratios remained stable with passages. CD271 and Stro-1 were expressed by <1% of the expanded cell populations. Interestingly, the percentage of CD56+cells strongly increased from P1 (25%) to P4 (80%) both in all sub-populations studied. CD146+CD56+, MSCA-1+CD56+, and CD146+MSCA-1+cells were the most abundant DP-MSCs at the end of P4. These results established that DP-MSCs constitute a heterogeneous mixture of cells in pulp tissuein vivoand in culture, and that their phenotype is modified uponin vitroexpansion. Further studies are needed to determine whether co-expression of specific MSC markers confers DP cells specific properties that could be used for the regeneration of human tissues, including the dental pulp, with standardized cell-based medicinal products.
authors.affiliation: Laboratoire de Biologie Tissulaire et Ingénierie Thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Lyon 1, UMS3444 BioSciences Gerland-Lyon SudLyon, France; Faculté d'Odontologie, Université de Lyon, Université Lyon 1Lyon, France; Hospices Civils de Lyon, Service d'OdontologieLyon, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Ducret
authors.affiliation: Laboratoire de Biologie Tissulaire et Ingénierie Thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Lyon 1, UMS3444 BioSciences Gerland-Lyon SudLyon, France; Laboratory of Regenerative Technologies, Department of Biomedical Engineering, University of BaselBasel, Switzerland.
authors.forename: Hugo
authors.initials: H
authors.lastname: Fabre
authors.affiliation: CTI-BIOTECH, Cell Therapy Research Institute Meyzieu, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Degoul
authors.affiliation: CTI-BIOTECH, Cell Therapy Research Institute Meyzieu, France.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Atzeni
authors.affiliation: CTI-BIOTECH, Cell Therapy Research Institute Meyzieu, France.
authors.forename: Colin
authors.initials: C
authors.lastname: McGuckin
authors.affiliation: CTI-BIOTECH, Cell Therapy Research Institute Meyzieu, France.
authors.forename: Nico
authors.initials: N
authors.lastname: Forraz
authors.affiliation: Laboratoire de Biologie Tissulaire et Ingénierie Thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Lyon 1, UMS3444 BioSciences Gerland-Lyon Sud Lyon, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Mallein-Gerin
authors.affiliation: Laboratoire de Biologie Tissulaire et Ingénierie Thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Lyon 1, UMS3444 BioSciences Gerland-Lyon Sud Lyon, France.
authors.forename: Emeline
authors.initials: E
authors.lastname: Perrier-Groult
authors.affiliation: Institut National De La Santé Et De La Recherche Médicale UMR1064, Faculté d'Odontologie, Centre de Recherche en Transplantation et Immunologie, Université de Nantes Nantes, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Alliot-Licht
authors.affiliation: Laboratoire de Biologie Tissulaire et Ingénierie Thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Lyon 1, UMS3444 BioSciences Gerland-Lyon SudLyon, France; Faculté d'Odontologie, Université de Lyon, Université Lyon 1Lyon, France; Hospices Civils de Lyon, Service d'OdontologieLyon, France.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Farges
keywords: CD146; CD56; cell-based medicinal products; human dental pulp; immunophenotyping; mesenchymal stromal cells; regenerative dentistry; stem cell markers