“…9 ). The novel methods and concepts presented here are relevant to the study of whole nerve tissues 41 , SC-like cells generated from stem cell sources 42 and neural crest-derived cells including but not restricted to SCs. Figure 9 Auto and heterotypic interactions between SCs and fibroblasts in nerve regeneration.…”
Section: Discussionmentioning
confidence: 99%
“…The unique potential of SCs for remyelination along with their provision of an extracellular environment supportive of neural regeneration have converted these glial cells into excellent candidates for transplantation in the PNS and CNS when used as purified cells or tissue grafts 41 , 50 . The traditional view that fibroblasts contribute scar tissue and negatively affect the outcome of SC transplantation is changing upon new evidence indicating that they assist in tissue regeneration.…”
Nerve-derived human Schwann cell (SC) cultures are irreplaceable models for basic and translational research but their use can be limited due to the risk of fibroblast overgrowth. Fibroblasts are an ill-defined population consisting of highly proliferative cells that, contrary to human SCs, do not undergo senescence in culture. We initiated this study by performing an exhaustive immunological and functional characterization of adult nerve-derived human SCs and fibroblasts to reveal their properties and optimize a protocol of magnetic-activated cell sorting (MACS) to separate them effectively both as viable and biologically competent cells. We next used immunofluorescence microscopy imaging, flow cytometry analysis and next generation RNA sequencing (RNA-seq) to unambiguously characterize the post-MACS cell products. High resolution transcriptome profiling revealed the identity of key lineage-specific transcripts and the clearly distinct neural crest and mesenchymal origin of human SCs and fibroblasts, respectively. Our analysis underscored a progenitor- or stem cell-like molecular phenotype in SCs and fibroblasts and the heterogeneity of the fibroblast populations. In addition, pathway analysis of RNA-seq data highlighted putative bidirectional networks of fibroblast-to-SC signaling that predict a complementary, yet seemingly independent contribution of SCs and fibroblasts to nerve regeneration. In sum, combining MACS with immunochemical and transcriptomics approaches provides an ideal workflow to exhaustively assess the identity, the stage of differentiation and functional features of highly purified cells from human peripheral nerve tissues.
“…9 ). The novel methods and concepts presented here are relevant to the study of whole nerve tissues 41 , SC-like cells generated from stem cell sources 42 and neural crest-derived cells including but not restricted to SCs. Figure 9 Auto and heterotypic interactions between SCs and fibroblasts in nerve regeneration.…”
Section: Discussionmentioning
confidence: 99%
“…The unique potential of SCs for remyelination along with their provision of an extracellular environment supportive of neural regeneration have converted these glial cells into excellent candidates for transplantation in the PNS and CNS when used as purified cells or tissue grafts 41 , 50 . The traditional view that fibroblasts contribute scar tissue and negatively affect the outcome of SC transplantation is changing upon new evidence indicating that they assist in tissue regeneration.…”
Nerve-derived human Schwann cell (SC) cultures are irreplaceable models for basic and translational research but their use can be limited due to the risk of fibroblast overgrowth. Fibroblasts are an ill-defined population consisting of highly proliferative cells that, contrary to human SCs, do not undergo senescence in culture. We initiated this study by performing an exhaustive immunological and functional characterization of adult nerve-derived human SCs and fibroblasts to reveal their properties and optimize a protocol of magnetic-activated cell sorting (MACS) to separate them effectively both as viable and biologically competent cells. We next used immunofluorescence microscopy imaging, flow cytometry analysis and next generation RNA sequencing (RNA-seq) to unambiguously characterize the post-MACS cell products. High resolution transcriptome profiling revealed the identity of key lineage-specific transcripts and the clearly distinct neural crest and mesenchymal origin of human SCs and fibroblasts, respectively. Our analysis underscored a progenitor- or stem cell-like molecular phenotype in SCs and fibroblasts and the heterogeneity of the fibroblast populations. In addition, pathway analysis of RNA-seq data highlighted putative bidirectional networks of fibroblast-to-SC signaling that predict a complementary, yet seemingly independent contribution of SCs and fibroblasts to nerve regeneration. In sum, combining MACS with immunochemical and transcriptomics approaches provides an ideal workflow to exhaustively assess the identity, the stage of differentiation and functional features of highly purified cells from human peripheral nerve tissues.
“…Nevertheless, clinical translation of human SC transplants was accomplished only recently. Whereas no major safety concerns have been raised so far, the efficacy of SC transplantation in the brain, spinal cord and nerve remains to be confirmed [recently reviewed in [8,11]).…”
Section: Cell Therapymentioning
confidence: 99%
“…Notoriously, it has been shown that SCs from cadaveric human nerves not only survive for a long time under cold storage but also remain potent to generate expandable SC cultures if placed under appropriate in vitro conditions [6,7]. These advantages have provided researchers with an opportunity to transplant isolated SCs or nerve tissue grafts to foster neuroprotection, neuroregeneration and remyelination after trauma or disease (reviewed in [8][9][10]).…”
Section: Introductionmentioning
confidence: 99%
“…Current protocols for SC culturing have mostly used the largest nerve in the body, the sciatic nerve, because of practical rather than biological reasons. For instance, clinical protocols for human SC culture have used the sural nerve, a sensory branch from the sciatic nerve in the leg, because its surgical removal can be relatively well tolerated by the patient [8,9]. Cultured SCs display unambiguous features regardless of the nerve of origin and the age of the donor though SCs from immature nerves (or younger donors) have a tendency to proliferate at a faster rate [6].…”
Schwann cell (SC) cultures from experimental animals and human donors can be prepared using nearly any type of nerve at any stage of maturation to render stage- and patient-specific populations. Methods to isolate, purify, expand in number, and differentiate SCs from adult, postnatal and embryonic sources are efficient and reproducible as these have resulted from accumulated refinements introduced over many decades of work. Albeit some exceptions, SCs can be passaged extensively while maintaining their normal proliferation and differentiation controls. Due to their lineage commitment and strong resistance to tumorigenic transformation, SCs are safe for use in therapeutic approaches in the peripheral and central nervous systems. This review summarizes the evolution of work that led to the robust technologies used today in SC culturing along with the main features of the primary and expanded SCs that make them irreplaceable models to understand SC biology in health and disease. Traditional and emerging approaches in SC culture are discussed in light of their prospective applications. Lastly, some basic assumptions in vitro SC models are identified in an attempt to uncover the combined value of old and new trends in culture protocols and the cellular products that are derived.
The work here reported analyzes the effect of increased efficiency of brain‐derived neurotrophic factor (BDNF) production by electroporated Schwann cells (SCs) on the axonal extension in a coculture system on a biomaterial platform that can be of interest for the treatment of injuries of the nervous system, both central and peripheral. Rat SCs are electrotransfected with a plasmid coding for the BDNF protein in order to achieve an increased expression and release of this protein into the culture medium of the cells, performing the best balance between the level of transfection and the number of living cells. Gene‐transfected SCs show an about 100‐fold increase in the release of BDNF into the culture medium, compared to nonelectroporated SCs. Cocultivation of electroporated SCs with rat dorsal root ganglia (DRG) is performed on highly aligned substrates of polylactic acid (PLA) microfibers coated with the electroconductive polymer polypyrrol (PPy). The coculture of DRG with electrotransfected SCs increase both the axonal extension and the axonal sprouting from DRG neurons compared to the coculture of DRG with nonelectroporated SCs. Therefore, the use of PLA–PPy highly aligned microfiber substrates preseeded with electrotransfected SCs with an increased BDNF secretion is capable of both guiding and accelerating axonal growth.
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