An allogeneic \u2018off the shelf\u2019 therapeutic strategy for peripheral nerve tissue engineering using clinical grade human neural stem cells

C O’Rourke,J B Phillips,A G E Day,L Stevanato,J Sinden,J Cowan,L Thanabalasundaram,C Murray-Dunning,N Grace,R A L Drake,G Cameron

doi:10.1038/s41598-018-20927-8

Abstract

Artificial tissues constructed from therapeutic cells offer a promising approach for improving the treatment of severe peripheral nerve injuries. In this study the effectiveness of using CTX0E03, a conditionally immortalised human neural stem cell line, as a source of allogeneic cells for constructing living artificial nerve repair tissue was tested. CTX0E03 cells were differentiated then combined with collagen to form engineered neural tissue (EngNT-CTX), stable aligned sheets of cellular hydrogel. EngNT-CTX sheets were delivered within collagen tubes to repair a 12 mm sciatic nerve injury model in athymic nude rats. Autologous nerve grafts (autografts) and empty tubes were used for comparison. After 8 weeks functional repair was assessed using electrophysiology. Further, detailed histological and electron microscopic analysis of the repaired nerves was performed. Results indicated that EngNT-CTX supported growth of neurites and vasculature through the injury site and facilitated reinnervation of the target muscle. These findings indicate for the first time that a clinically validated allogeneic neural stem cell line can be used to construct EngNT. This provides a potential ‘off the shelf’ tissue engineering solution for the treatment of nerve injury, overcoming the limitations associated with nerve autografts or the reliance on autologous cells for populating repair constructs.

Highlights

Tissue engineering provides opportunities to combine therapeutic cells with materials in order to construct living artificial tissues that can repair nervous system injury where significant amounts of tissue have been lost
TM stabilisation to form engineered neural tissue (EngNT)-CTX, which was rolled into rods and placed within NeuraGen conduits for testing in vivo using athymic nude rats
In one of the EngNT samples and TM TM two of the NeuraGen samples Compound muscle action potentials (CMAP) amplitude was below the threshold of detection

Summary

Introduction

Tissue engineering provides opportunities to combine therapeutic cells with materials in order to construct living artificial tissues that can repair nervous system injury where significant amounts of tissue have been lost. In order for this approach to be successful it is necessary to identify sources of cells that are compatible with tissue engineering technology and suitable for translation to the clinic and eventually commercial manufacture. The proven suitability for clinical translation of CTX0E03 combined with its nervous system origin make CTX0E03 a promising candidate cell line for use in neural tissue engineering, for the treatment of peripheral nerve injuries where there are currently no living allogeneic cellular repair options available. The key difference between the autograft and these other approaches is the presence of columns of living Schwann cells that are present in the autograft and provide support and guidance to regenerating neurons[15]

Methods

Results

Conclusion