Towards the development of a pragmatic technique for isolating and differentiating nestin‐positive cells from human scalp skin into neuronal and glial cell populations: generating neurons from human skin?

Abstract

Nestin+ hair follicle-associated cells of murine skin can be isolated and differentiated in vitro into neuronal and glial cells. Therefore, we have asked whether human skin also contains nestin+ cells, and whether these can be differentiated in vitro into neuronal and/or glial cell populations. In this methodological pilot study, we show that both are indeed the case - employing purposely only very simple techniques for isolating, propagating, and differentiating nestin+ cells from normal human scalp skin and its appendages that do not require selective microdissection and tissue compartment isolation prior to cell culture. We show that, it is in principle, possible to maintain and propagate human skin nestin+ cells for extended passage numbers and to differentiate them into both neuronal (i.e. neurofilament+ and/or PGP9.5+) and glial (i.e. GFAP+, MBP+ and/or O4+) cell populations. Therefore, human scalp skin can serve as a highly accessible, abundant, and convenient source for autologous adult stem cell-like cells that offer themselves to be exploited for neuroregenerative medicine purposes.

Spatiotemporal expression pattern of neuroepithelial stem cell marker nestin suggests a role in dermal homeostasis, neovasculogenesis, and tumor stroma development: A study on embryonic and adult human skin

“…[42][43][44] From the literature quoted so far and from our own results it is apparent that nestin expression apart from activated melanocytes is confined to mesodermally derived cells (CTS and endothelial cells). Our results are supported by a study from Kruse et al 45 Staining pattern for nestin is identical with 3 different nestin antibodies. All 3 nestin antibodies display identical staining pattern in regard to all anatomic structures of all different tissue samples examined.…”

“…Original magnifications: A to C, Hair germ stage, 14 weeks (3400); D to F, bulbous hair peg stage, 19 weeks (3100); G to I, lower stem and hair bulb of postnatal hair follicle (3200); and J to L, BCC (3200). m J AM ACAD DERMATOL VOLUME 63, NUMBER 1 et al 46 We suspect technical differences in the procedure used by these authors for this discrepancy in the immunohistochemical findings between our results and those of Kruse et al 45 46 We used the same mouse monoclonal antibody from Santa Cruz Biotechnology as Wang et al 46 and confirmed our results different from theirs with two other antibodies.…”

“…51,54,55 In an extension of their original studies on the neuronal differentiation supposedly of the bulge keratinocytes in the murine model the authors also published a series of articles maintaining that nestin-positive follicle bulge cells are also capable of differentiating into blood vessels. [56][57][58] The localization of nestin-positive cells within the epithelial component of the hair follicle is a major departure from the observations by Kruse et al 45 and our own, because we both located the nestin-positive cells in the mesenchymal compartments of the skin. In fact, we specifically commented that with all 3 nestin antibodies the bulge is negative for nestin-immunoreactive cells with the exception of incidental melanocytes.…”

“…However, our results are not agreement with a study by Wang et al46 reporting all layers of the epidermis as nestin-positive, with basal cells being more immunoreactive than those in the superbasal layer. Because Kruse et al45 and Wang et al 46 both used frozen sections (the first group acetone fixed, the second one paraformaldehyde fixed), it is unlikely that the use of frozen sections is the reason for this discrepancy between the results. In a most recent publication by Kanoh et al 47 nestin was not detected in the epidermis.…”

Adult epidermal Notch activity induces dermal accumulation of T cells and neural crest derivatives through upregulation of jagged 1

“…The intermediate filament protein nestin is expressed by peripheral nerves adjacent to the hair follicle bulge and by neural crest stem cells, melanocyte precursors and dermal papilla cells (Amoh et al, 2005;Dunn et al, 2000;Kruse et al, 2006;Li et al, 2003;Tiede et al, 2007). In K14NICDER mice treated with 4OHT for 14 days, there was marked accumulation of nestin-positive cells at the epidermal-dermal boundary (Fig.…”