title: Self-renewal and multilineage differentiation of mouse dental epithelial stem cells.
abstract: Understanding the cellular and molecular mechanisms underlying the self-renewal and differentiation of dental epithelial stem cells (DESCs) that support the unlimited growth potential of mouse incisors is critical for developing novel tooth regenerative therapies and unraveling the pathogenesis of odontogenic tumors. However, analysis of DESC properties and regulation has been limited by the lack of an in vitro assay system and well-documented DESC markers. Here, we describe an in vitro sphere culture system to isolate the DESCs from postnatal mouse incisor cervical loops (CLs) where the DESCs are thought to reside. The dissociated cells from CLs were able to expand and form spheres for multiple generations in the culture system. Lineage tracing indicated that DESC within the spheres were epithelial in origin as evident by lineage tracing. Upon stimulation, the sphere cells differentiated into cytokeratin 14- and amelogenin-expressing and mineral material-producing cells. Compared to the CL tissue, sphere cells expressed high levels of expression of Sca-1, CD49f (also designated as integrin α6), and CD44. Fluorescence-activated cell sorting (FACS) analyses of mouse incisor CL cells further showed that the CD49f(Bright) population was enriched in sphere-forming cells. In addition, the CD49f(Bright) population includes both slow-cycling and Lgr5(+) DESCs. The in vitro sphere culture system and identification of CD49f(Bright) as a DESC marker provide a novel platform for enriching DESCs, interrogating how maintenance, cell fate determination, and differentiation of DESCs are regulated, and developing tooth regenerative therapies.
authors.affiliation: Center for Cancer and Stem Cell Biology, Institute of Biosciences and Technology, Texas A&M University, Houston, TX 77030-3303, USA; Department of Biomedical Sciences, Baylor College of Dentistry, Texas A&M University, Houston, TX 77030-3303, USA. Electronic address: jyfchang@uw.edu.
authors.forename: Julia Yu Fong
authors.initials: JY
authors.lastname: Chang
authors.forename: Cong
authors.initials: C
authors.lastname: Wang
authors.forename: Chengliu
authors.initials: C
authors.lastname: Jin
authors.forename: Chaofeng
authors.initials: C
authors.lastname: Yang
authors.forename: Yanqing
authors.initials: Y
authors.lastname: Huang
authors.forename: Junchen
authors.initials: J
authors.lastname: Liu
authors.forename: Wallace L
authors.initials: WL
authors.lastname: McKeehan
authors.forename: Rena N
authors.initials: RN
authors.lastname: D'Souza
authors.forename: Fen
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D000950:Antigens, Ly / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism; D007180:Incisor / Q000166:cytology* / Q000378:metabolism; D039503:Integrin alpha6 / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D000950:Antigens, Ly; D015415:Biomarkers; D018960:Hyaluronan Receptors; D039503:Integrin alpha6; C493597:Lgr5 protein, mouse; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D043562:Receptors, G-Protein-Coupled
keywords: Alp; CL; DESC; FGF; FGFR; IEE; OEE; SC; SI; SR; alkaline phosphatase; cervical loop; dental epithelial stem cell; fibroblast growth factor; fibroblast growth factor receptor; inner enamel epithelium; outer enamel epithelium; stellate reticulum; stem cell; stratum intermedium
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