title: α6 Integrin (α6high)/Transferrin Receptor (CD71)lowKeratinocyte Stem Cells Are More Potent for Generating Reconstructed Skin Epidermis Than Rapid Adherent Cells.
abstract: The epidermis basal layer is composed of two keratinocyte populations: Keratinocyte Stem cells (KSC) and Transitory Amplifying (TA) cells that arise from KSC division. Unfortunately, no specific marker exists to differ between KSC and TA cells. Here, we aimed at comparing two different methods that pretended to isolate these two populations: (i) the rapid adhesion method on coated substrate and (ii) the flow cytometry method, which is based on the difference in cell surface expressions of the α6 integrin and transferrin receptor (CD71). Then, we compared different parameters that are known to discriminate KSC and TA populations. Interestingly, we showed that both methods allow enrichment in stem cells. However, cell sorting by flow cytometry (α6high/CD71low) phenotype leads to a better enrichment of KSC since the colony forming efficiency is five times increased versus total cell suspension, whereas it is only 1.4 times for the adhesion method. Moreover, α6high/CD71lowcells give rise to a thicker pluristratified epithelium with lower seeding density and display a low Ki67 positive cells number, showing that they have reached the balance between proliferation and differentiation. We clearly demonstrated that cells isolated by a rapid adherent method are not the same population as KSC isolated by flow cytometry following α6high/CD71lowphenotype.
authors.affiliation: Gattefossé, 36 chemin de Genas, F-69800 Saint-Priest, France. odile.damour@chu-lyon.fr.|Commissariat à l'énergie atomique et aux énergies alternatives (CEA)/Institut Nanosciences et cryogénie (INAC)/SYstèmes Moléculaires et nanoMatériaux pour l'Energie et la Santé (SyMMES)/Lésions des acides nucléiques (LAN), 17 avenue des martyrs, F-38054 Grenoble CEDEX, France. odile.damour@chu-lyon.fr.|Hospices Civils de LYON (HCL)/Banque de Tissus et Cellules/Laboratoire des Substituts Cutanés, 5 place d'Arsonval, F-69000 Lyon, France. odile.damour@chu-lyon.fr.|Department of Biological Sciences, University Grenoble Alpes, F-38000 Grenoble, France. odile.damour@chu-lyon.fr.
authors.forename: Elodie
authors.initials: E
authors.lastname: Metral
authors.affiliation: Gattefossé, 36 chemin de Genas, F-69800 Saint-Priest, France. nbechetoille@gattefosse.com.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bechetoille
authors.affiliation: Gattefossé, 36 chemin de Genas, F-69800 Saint-Priest, France. fdemarne@gattefosse.com.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Demarne
authors.affiliation: Commissariat à l'énergie atomique et aux énergies alternatives (CEA)/Institut Nanosciences et cryogénie (INAC)/SYstèmes Moléculaires et nanoMatériaux pour l'Energie et la Santé (SyMMES)/Lésions des acides nucléiques (LAN), 17 avenue des martyrs, F-38054 Grenoble CEDEX, France. walid.rachidi@cea.fr.|Department of Biological Sciences, University Grenoble Alpes, F-38000 Grenoble, France. walid.rachidi@cea.fr.
authors.forename: Walid
authors.initials: W
authors.lastname: Rachidi
authors.affiliation: Hospices Civils de LYON (HCL)/Banque de Tissus et Cellules/Laboratoire des Substituts Cutanés, 5 place d'Arsonval, F-69000 Lyon, France. odile.damour@chu-lyon.fr.
authors.forename: Odile
authors.initials: O
authors.lastname: Damour
mesh_terms: D015703:Antigens, CD / Q000378:metabolism*; D002448:Cell Adhesion; D002469:Cell Separation; D002999:Clone Cells; D024042:Collagen Type I / Q000378:metabolism; D003114:Colony-Forming Units Assay; D000078404:Epidermal Cells; D005434:Flow Cytometry; D006801:Humans; D039503:Integrin alpha6 / Q000378:metabolism*; D015603:Keratinocytes / Q000166:cytology*; D010641:Phenotype; D011990:Receptors, Transferrin / Q000378:metabolism*; D012038:Regeneration; D013234:Stem Cells / Q000378:metabolism*; D013696:Temperature
chemical_list: D015703:Antigens, CD; C068126:CD71 antigen; D024042:Collagen Type I; D039503:Integrin alpha6; D011990:Receptors, Transferrin
keywords: CD71; adhesion; basal keratinocytes; epidermis; keratinocyte stem cells; skin; α6 integrin