title: Characterization of neurons from adult human skin-derived precursors in serum-free medium : a PCR array and immunocytological analysis.
abstract: Adult stem cells could be small sources of neurons or other cellular types for regenerative medicine and tissue engineering. Recently, pluripotent stem cells have been extracted from skin tissue, which opened a new accessible source for research. To routinely obtain a high yield of functional neurons from adult human skin stem cells with defined serum-free medium, stem cells from abdominal skin were cultured in serum-free medium. To differentiate them, we used a defined medium containing growth factors. Differentiated cells were identified using the following methods: (i) Oil-red-O staining for adipocytes, immunocytochemistry with antibodies recognising (ii) neurofilaments and PGP9.5 for neural differentiation, (iii) glial fibrillary acidic protein (GFAP) for glial differentiation, (iv) Ki-67 for proliferative cells, (v) FM1-43 staining to analyse vesicle trafficking in neuronal cells and (vi) a PCR array was used. Stem cells were floating in spheres and were maintained in culture for 4 months or more. They expressed nestin and Oct 4 and were proliferative. We induced specific differentiation into adipocytes, glial and neuronal cells. The yields of differentiated neurons were high and reproducible. They were maintained for long time (1 month) in the culture medium. Furthermore, these neurons incorporated FM1-43 dye, which indicates a potent acquisition of synaptic features in neurons. Stem cells from adult human skin could be valuable and reproducible tools/source to obtain high numbers of functional specific cellular types, such as neurons, for tissue engineering. In this work, the possibility to obtain a high yield of differentiated neurons, with the ability of endocytosis and vesicle cell trafficking, was shown.
authors.affiliation: Faculty of Medicine and Health Sciences, Laboratory of Skin Neurobiology, University of Brest, Brest, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Lebonvallet
authors.forename: Nicholas
authors.initials: N
authors.lastname: Boulais
authors.forename: Christelle
authors.initials: C
authors.lastname: Le Gall
authors.forename: Jeremy
authors.initials: J
authors.lastname: Chéret
authors.forename: Ulysse
authors.initials: U
authors.lastname: Pereira
authors.forename: Olivier
authors.initials: O
authors.lastname: Mignen
authors.forename: Vincent
authors.initials: V
authors.lastname: Bardey
authors.forename: Christine
authors.initials: C
authors.lastname: Jeanmaire
authors.forename: Louis
authors.initials: L
authors.lastname: Danoux
authors.forename: Gilles
authors.initials: G
authors.lastname: Pauly
authors.forename: Laurent
authors.initials: L
authors.lastname: Misery
mesh_terms: D017667:Adipocytes / Q000166:cytology; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology; D016895:Culture Media, Serum-Free; D004705:Endocytosis / Q000502:physiology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D058953:Neural Stem Cells / Q000502:physiology*; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology*; D016133:Polymerase Chain Reaction; D012867:Skin / Q000166:cytology*; D013572:Synaptic Vesicles / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free