title: EGF-FGF2 stimulates the proliferation and improves the neuronal commitment of mouse epidermal neural crest stem cells (EPI-NCSCs).
abstract: Epidermal neural crest stem cells (EPI-NCSCs), which reside in the bulge of hair follicles, are attractive candidates for several applications in cell therapy, drug screening and tissue engineering. As suggested remnants of the embryonic neural crest (NC) in an adult location, EPI-NCSCs are able to generate a wide variety of cell types and are readily accessible by a minimally invasive procedure. Since the combination of epidermal growth factor (EGF) and fibroblast growth factor type 2 (FGF2) is mitogenic and promotes the neuronal commitment of various stem cell populations, we examined its effects in the proliferation and neuronal potential of mouse EPI-NCSCs. By using a recognized culture protocol of bulge whiskers follicles, we were able to isolate a population of EPI-NCSCs, characterized by the migratory potential, cell morphology and expression of phenotypic markers of NC cells. EPI-NCSCs expressed neuronal, glial and smooth muscle markers and exhibited the NC-like fibroblastic morphology. The treatment with the combination EGF and FGF2, however, increased their proliferation rate and promoted the acquisition of a neuronal-like morphology accompanied by reorganization of neural cytoskeletal proteins βIII-tubulin and nestin, as well as upregulation of the pan neuronal marker βIII-tubulin and down regulation of the undifferentiated NC, glial and smooth muscle cell markers. Moreover, the treatment enhanced the response of EPI-NCSCs to neurogenic stimulation, as evidenced by induction of GAP43, and increased expression of Mash-1 in neuron-like cell, both neuronal-specific proteins. Together, the results suggest that the combination of EGF-FGF2 stimulates the proliferation and improves the neuronal potential of EPI-NCSCs similarly to embryonic NC cells, ES cells and neural progenitor/stem cells of the central nervous system and highlights the advantage of using EGF-FGF2 in neuronal differentiation protocols.
authors.affiliation: Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Raul Bardini
authors.initials: RB
authors.lastname: Bressan
authors.affiliation: Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Fernanda Rosene
authors.initials: FR
authors.lastname: Melo
authors.affiliation: Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Patricia Alves
authors.initials: PA
authors.lastname: Almeida
authors.affiliation: Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Denise Avani
authors.initials: DA
authors.lastname: Bittencourt
authors.affiliation: Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Silvia
authors.initials: S
authors.lastname: Visoni
authors.affiliation: Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Talita Silva
authors.initials: TS
authors.lastname: Jeremias
authors.affiliation: Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Costa
authors.affiliation: Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil.
authors.forename: Rodrigo Bainy
authors.initials: RB
authors.lastname: Leal
authors.affiliation: Departamento de Biologia Celular, Embriologia e Genética, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário - Trindade, 88040-900 Florianópolis SC, Brazil. Electronic address: andrea.trentin@ufsc.br.
authors.forename: Andrea Gonçalves
authors.initials: AG
authors.lastname: Trentin
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D015536:Down-Regulation / Q000502:physiology; D004815:Epidermal Growth Factor / Q000378:metabolism*; D004817:Epidermis / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D019922:GAP-43 Protein / Q000378:metabolism; D018859:Hair Follicle / Q000378:metabolism; D051379:Mice; D039902:Multipotent Stem Cells / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D009432:Neural Crest / Q000378:metabolism*; D058953:Neural Stem Cells / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D004815:Epidermal Growth Factor; D016222:Fibroblast Growth Factor 2; D019922:GAP-43 Protein; C499317:Ascl1 protein, mouse
keywords: Cell differentiation; Cell fate; Cell proliferation; Cell therapy; Neural crest; Neural progenitors; Neurogenesis