Self-aligned Schwann cell monolayers demonstrate an inherent ability to direct neurite outgrowth

Abstract

In vivo nerve guidance channel studies have identified Schwann cell (SC) presence as an integral factor in axonal number and extension in an injury site, and in vitro studies have provided evidence that oriented SCs can direct neurite outgrowth. However, traditional methods used to create oriented SC monolayers (e.g. micropatterns/microtopography) potentially introduce secondary guidance cues to the neurons that are difficult to de-couple. Although SCs expanded on uniform laminin-coated coverslips lack a global orientation, the monolayers contain naturally formed regions of locally oriented cells that can be used to investigate SC-mediated neurite guidance. In this work, novel image analysis techniques have been developed to quantitatively assess local neurite orientation with respect to the underlying regional orientation of the Schwann cell monolayer. Results confirm that, in the absence of any secondary guidance cues, a positive correlation exists between neurite outgrowth and regional orientation of the SC monolayer. Thus, SCs alone possess an inherent ability to direct neurite outgrowth, and expansion of the co-culture-based quantitative method described can be used to further deconstruct specific biomolecular mechanisms of neurite guidance.

A new three-dimensional axonal outgrowth assay for central nervous system regeneration

“…5). Neurite outgrowth has been reported to be directed by Schwann cell alignment in plane culture (Seggio et al, 2010;Thompson and Buettner, 2006). However, Schwann cells in this 3D culture system were aligned to the regenerating axons, which is quite different from the situation observed for 2D culture.…”

Optogenetic neuronal stimulation promotes axon outgrowth and myelination of motor neurons in a three‐dimensional motor neuron–Schwann cell coculture model on a microfluidic biochip

“…Methods including the use of neurotrophic factors (Meyer, Matsuoka, Wetmore, Olson, & Thoenen, 1992;Sendtner, Kreutzberg, & Thoenen, 1990), supportive cell grafts (e.g., SCs; Seggio, Narayanaswamy, Roysam, & Thompson, 2010) or stem cells (Zhang et al, 2010), autografts (den Dunnen & Meek, 2001), and artificial nerve conduits (Nectow, Marra, & Kaplan, 2012;Xu et al, 2003) have been shown to influence axon outgrowth and myelination both in vivo and in vitro. Methods including the use of neurotrophic factors (Meyer, Matsuoka, Wetmore, Olson, & Thoenen, 1992;Sendtner, Kreutzberg, & Thoenen, 1990), supportive cell grafts (e.g., SCs; Seggio, Narayanaswamy, Roysam, & Thompson, 2010) or stem cells (Zhang et al, 2010), autografts (den Dunnen & Meek, 2001), and artificial nerve conduits (Nectow, Marra, & Kaplan, 2012;Xu et al, 2003) have been shown to influence axon outgrowth and myelination both in vivo and in vitro.…”

“…Numerous strategies for promoting axon regrowth and remyelination following injury have been investigated. Methods including the use of neurotrophic factors (Meyer, Matsuoka, Wetmore, Olson, & Thoenen, 1992;Sendtner, Kreutzberg, & Thoenen, 1990), supportive cell grafts (e.g., SCs; Seggio, Narayanaswamy, Roysam, & Thompson, 2010) or stem cells (Zhang et al, 2010), autografts (den Dunnen & Meek, 2001), and artificial nerve conduits (Nectow, Marra, & Kaplan, 2012;Xu et al, 2003) have been shown to influence axon outgrowth and myelination both in vivo and in vitro. In addition, electrical stimulation (ES) has long been considered a potential method to enhance recovery in both the central nervous system (CNS) and the PNS (McCaig & Rajnicek, 1991;Thompson, Koppes, Hardy, & Schmidt, 2014).…”

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An <em>In Vitro</em> Co-culture Model to Study Peripheral Nerve Regeneration

“…The method proposed in this paper is a revised version of a well-established protocol in our laboratory 11 and it involves a direct contact between the two cell types (direct co-culture), in which the second cell type (DRG neurons) are seeded on the top of the others (SC-like ASC). This approach is based on previous findings that demonstrated the importance of the presence of a glial cell layer on the substrate when seeding neuronal cultures [35][36][37] . This effect is likely due to cell-cell interactions through DRG integrins and ECM molecules deposited from ASC and other cues on the stem cell surface.…”