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.

Biotechnological Management of Skin Burn Injuries: Challenges and Perspectives in Wound Healing and Sensory Recovery

“…Neuronal differentiation is achieved using AMPc and a cocktail of neurotrophins such as BDNF, NT-3 and NGF while glial differentiation into Schwann cell is promoted by the addition of forskolin and heregulin 1 to the culture medium(267)(268)(269)(270) (Figure 6). Little is known regarding the role of SKPs in skin wound healing or a potential involvement in sensory nerve regrowth but several studies have shown that SKP-derived Schwann cells help promoting sciatic nerve regeneration in rodents(271)(272)(273).…”

New insights into the roles of myofibroblasts and innervation during skin healing and innovative therapies to improve scar innervation

“…Strategies using neuropeptides to stimulate skin cells and trophic factors to stimulate neurite growth can be either combined or provided sequentially. A possible source of human neuropeptides, ideally from sensory neurones, could be generated using skin‐derived precursors (SKPs) or induced pluripotent stem cells (iPS) differentiated into neurones . Due to the distinct role of different neuropeptides and NTs, the use of a cocktail would be interesting with delivery either by single administration or ideally by delayed delivery into the wound .…”

Induction of Schwann Cells from Rat Bone Marrow Mesenchymal Stem Cells

“…In addition, tumorigenicity of ES cells and iPS cells is another concern in application to cell therapy. Recent studies have elucidated the existence of a particular type of stem cells such as skin-derived precursors (SKPs) in the adult human dermis, which exhibit the similar properties to neural crest cells (Gingras 2007;Joannides 2004;Lebonvallet 2012). SKPs are reported to be able to differentiate into Schwann cells within a week in a high rate in vitro (>95%) (McKenzie 2006), suggesting that SKPs will be a good candidate as a source for Schwann cell generation.…”