α6 Integrin (α6 high )/Transferrin Receptor (CD71) low Keratinocyte Stem Cells Are More Potent for Generating Reconstructed Skin Epidermis Than Rapid Adherent Cells

• Published in: International journal of molecular sciences Vol. 18; no. 2; p. 282
• Main Authors: Metral, Elodie, Bechetoille, Nicolas, Demarne, Frédéric, Rachidi, Walid, Damour, Odile
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 27.01.2017; MDPI
• Subjects: Adherent cells; Adhesion; Antigens, CD - metabolism; basal keratinocytes; Biochemistry, Molecular Biology; Biotechnology; Cancer; CD71; Cell Adhesion; Cell Behavior; Cell differentiation; Cell proliferation; Cell Separation; Cell surface; Cellular Biology; Clone Cells; Collagen Type I - metabolism; Colony-Forming Units Assay; Dermatology; Epidermis; Epidermis - cytology; Epithelium; Flow Cytometry; Genomics; Human health and pathology; Humans; Integrin alpha6 - metabolism; keratinocyte stem cells; Keratinocytes - cytology; Life Sciences; Molecular biology; Phenotype; Phenotypes; Populations; Receptors, Transferrin - metabolism; Regeneration; skin; Stem cells; Stem Cells - metabolism; Subcellular Processes; Temperature; Transferrin; Transferrins; α6 integrin; CD71; basal keratinocytes; skin; adhesion; keratinocyte stem cells; epidermis; α6 integrin
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms18020282

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 (α6 /CD71 ) 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, α6 /CD71 cells 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 α6 /CD71 phenotype.