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Phenotypic Identification of Dental Pulp Mesenchymal Stem/Stromal Cells Subpopulations with Multiparametric Flow Cytometry

  • Maxime Ducret
  • Jean-Christophe Farges
  • Marielle Pasdeloup
  • Emeline Perrier-Groult
  • Andreas Mueller
  • Frédéric Mallein-Gerin
  • Hugo Fabre
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1922)

Abstract

Dental pulp (DP) is a specialized, highly vascularized, and innervated connective tissue mainly composed of undifferentiated mesenchymal cells, fibroblasts, and highly differentiated dentin-forming odontoblasts. Undifferentiated mesenchymal cells include stem/stromal cell populations usually called dental pulp mesenchymal stem cells (DP-MSCs) which differ in their self-renewal properties, lineage commitment, and differentiation capabilities. Analysis of surface antigens has been largely used to precisely identify these DP-MSC populations. However, a major difficulty is that these antigens are actually not specific for MSCs. Most of the markers used are indeed shared by other cell populations such as progenitor cells, mature fibroblasts, and/or perivascular cells. Accordingly, the detection of only one of these markers in a cell population is clearly insufficient to determine its stemness. Recent data reported that multiparametric flow cytometry, by allowing for the detection of several molecules on the surface of one single cell, is a powerful tool to elucidate the phenotype of a cell population both in vivo and in vitro. So far, DP-MSC populations have been characterized mainly based on the isolated expression of molecules known to be expressed by stem cells, such as Stro-1 antigen, melanoma cell adhesion molecule MCAM/CD146, low-affinity nerve growth factor receptor p75NTR/CD271, and the mesenchymal stem cell antigen MSCA-1. Using multiparametric flow cytometry, we recently showed that human DP-MSCs are indeed phenotypically heterogeneous and form several populations.

The present paper describes the multiparametric flow cytometry protocol we routinely use for characterizing DP-MSCs. The description includes the design of the antibody panel and explains the selection of the different parameters related to the data quality control.

Key words

Dental pulp Mesenchymal stem/stromal cells Flow cytometry 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Maxime Ducret
    • 1
    • 2
    • 3
  • Jean-Christophe Farges
    • 1
    • 2
    • 3
  • Marielle Pasdeloup
    • 1
  • Emeline Perrier-Groult
    • 1
  • Andreas Mueller
    • 4
  • Frédéric Mallein-Gerin
    • 1
  • Hugo Fabre
    • 1
    • 4
  1. 1.Laboratoire de Biologie Tissulaire et Ingénierie ThérapeutiqueUMR5305 CNRS/Université Lyon 1, UMS3444 BioSciences Gerland-Lyon SudLyonFrance
  2. 2.Faculté d’Odontologie, Université de Lyon, Université Lyon 1LyonFrance
  3. 3.Hospices Civils de LyonService de Consultations et Traitements DentairesLyonFrance
  4. 4.Laboratory of Regenerative Technologies, Department of Biomedical EngineeringUniversity of BaselBaselSwitzerland

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