Aligned electrospun fibers for neural patterning

Erfan Soliman,Hua Ye,Fabio Bianchi,M Zameel Cader,Julian H George,James N Sleigh,Zhanfeng Cui

doi:10.1007/s10529-017-2494-z

Abstract

ObjectivesTo test a 3D approach for neural network formation, alignment, and patterning that is reproducible and sufficiently stable to allow for easy manipulation.ResultsA novel cell culture system was designed by engineering a method for the directional growth of neurons. This uses NG108-15 neuroblastoma x glioma hybrid cells cultured on suspended and aligned electrospun fibers. These fiber networks improved cellular directionality, with alignment angle standard deviations significantly lower on fibers than on regular culture surfaces. Morphological studies found nuclear aspect ratios and cell projection lengths to be unchanged, indicating that cells maintained neural morphology while growing on fibers and forming a 3D network. Furthermore, fibronectin-coated fibers enhanced neurite extensions for all investigated time points. Differentiated neurons exhibited significant increases in average neurite lengths 96 h post plating, and formed neurite extensions parallel to suspended fibers, as visualized through scanning electron microscopy.ConclusionsThe developed model has the potential to serve as the basis for advanced 3D studies, providing an original approach to neural network patterning and setting the groundwork for further investigations into functionality.

Highlights

Tissue-engineered platforms provide specialized mechanical and environmental cues in vitro for dissociated cells to grow, expand, and differentiate in conditions resembling in vivo settings
The developed model has the potential to serve as the basis for advanced 3D studies, providing an original approach to neural network patterning and setting the groundwork for further investigations into functionality
Differentiation was induced through serum starvation and the addition of dimethyl sulfoxide (DMSO) once cells were * 60% confluent

Summary

Introduction

Tissue-engineered platforms provide specialized mechanical and environmental cues in vitro for dissociated cells to grow, expand, and differentiate in conditions resembling in vivo settings. Such tissue environments can be invaluable for understanding cellular processes, functional development, and for more disease-relevant drug testing. Patterning of substrates on 2D surfaces can be used to provide guidance to growing neurons. Common techniques include micro-contact printing with chemo attractant protein inks (James et al 2000), micro-pillar construction for substrate patterning (Dowell-Mesfin et al 2004), aligned deposited fibers (Wu et al 2015) and coating with rough surfaces (Kim et al 2006). Neurons preferentially respond to environments patterned to better resemble in vivo conditions, even without biological cues (Richardson et al 2011)

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