Purification and Characterization of Schwann Cells from Adult Human Skin and Nerve

Abstract

Despite its modest capacity for regeneration, peripheral nervous system injury often results in significant long-term disability. Supplementing peripheral nervous system injury with autologous Schwann cells (SCs) may serve to rejuvenate the postinjury environment to enhance regeneration and ultimately improve functional outcomes. However, human nerve-derived SC (hN-SC) collection procedures require invasive surgical resection. Here, we describe the characterization of SCs from adult human skin (hSk-SCs) of four male donors ranging between 27 and 46 years old. Within five weeks of isolating and culturing adherent mixed skin cells, we were able to obtain 3–5 million purified SCs. We found that hSk-SCs appeared transcriptionally indistinguishable from hN-SCs with both populations exhibiting expression of SC genes including: SOX10, SOX9, AP2A1, CDH19, EGR1, ETV5, PAX3, SOX2, CX32, DHH, NECL4, NFATC4, POU3F1, S100B, and YY1. Phenotypic analysis of hSk-SCs and hN-SCs cultures revealed highly enriched populations of SCs indicated by the high percentage of NES+ve, SOX10+ve, s100+ve and p75+ve cells, as well as the expression of a battery of other SC-associated proteins (PAX3, CDH19, ETV5, SOX2, POU3F1, S100B, EGR2, and YY1). We further show that both hSk-SCs and hN-SCs are capable of promoting axonal growth to similar degrees and that a subset of both associate with regenerating axons and form myelin following transplantation into the injured mouse sciatic nerve. Interestingly, although the majority of both hSk-SCs and hN-SCs maintained SOX10 immunoreactivity following transplant, only a subset of each activated the promyelinating factor, POU3F1, and were able to myelinate. Taken together, we demonstrate that adult hSk-SCs are genetically and phenotypically indistinguishable to hN-SCs.

Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach

“…3 a). The transcription factors Sox2 and Oct6 exhibited SC-specific nuclear localization, as shown previously 26 , but their uneven levels of expression within the S100β positive population limited their use to discriminate SCs from fibroblasts effectively (not shown). Consistent with a previous report 27 , the intermediate filament vimentin and the anchoring membrane receptor CD44 were evenly expressed in human SCs and fibroblasts (Fig.…”

Etv5is not required for Schwann cell development but is required to regulate the Schwann cell response to peripheral nerve injury

“…Schwann cell transplants have the potential to aid peripheral nerve repair, and efforts are being made to improve the isolation and expansion of these cells to provide an adequate source for repair purposes. In this regard it is interesting that both human nervederived and skin derived Schwann cells cultured in vitro express a large number of Schwann cell markers associated with an early developmental phenotype, which includes Etv5 (Stratton et al, 2017). In our study here we provide the first glimpse that a decline in Etv5 expression leads to more number of Schwann cells following a peripheral nerve crush injury.…”

Novel insights into the glia limitans of the olfactory nervous system

“…The primary antibodies (Table ) used were: (1) goat anti‐GFAP (RRID: AB_880202) which has previously been used extensively by others to determine the activation state of astrocytes in CNS tissue and cells, see for example Ni et al (). (2) Rabbit anti‐S100β (RRID: AB_10013383), which has been used in many studies to label glial cells (in particular peripheral glia; both immunohistochemistry and immunocytochemistry), see for example Stratton et al (). We have previously used these two antibodies to characterize expression of GFAP and S100β in astrocytes in both tissue sections and cells (Chehrehasa et al, ; Windus et al, ).…”

“…Nuclei are stained with 4 0 ,6-diamidino-2-pheylindole in (c-f ). Scale bar is 290 μm in (c); 20 μm in (d, f, h-j); 440 μm in (e); 350 μm in (g); 180 μm in (k) [Color figure can be viewed at wileyonlinelibrary.com] cells (in particular peripheral glia; both immunohistochemistry and immunocytochemistry), see for example Stratton et al (2017). We have previously used these two antibodies to characterize expression of GFAP and S100β in astrocytes in both tissue sections and cells (Chehrehasa et al, 2010;Windus et al, 2010).…”