Elsevier

Neuroscience

Volume 164, Issue 3, 15 December 2009, Pages 1097-1107
Neuroscience

Clinical Neuroscience
Research Paper
Supplementation of acellular nerve grafts with skin derived precursor cells promotes peripheral nerve regeneration

https://doi.org/10.1016/j.neuroscience.2009.08.072Get rights and content

Abstract

Introduction of autologous stem cells into the site of a nerve injury presents a promising therapy to promote axonal regeneration and remyelination following peripheral nerve damage. Given their documented ability to differentiate into Schwann cells (SCs) in vitro, we hypothesized that skin-derived precursor cells (SKPs) could represent a clinically-relevant source of transplantable cells that would enhance nerve regeneration following peripheral nerve injury. In this study, we examined the potential for SKP-derived Schwann cells (SKP–SCs) or nerve-derived SCs to improve nerve regeneration across a 12 mm gap created in the sciatic nerve of Lewis rats bridged by a freeze-thawed nerve graft. Immunohistology after 4 weeks showed survival of both cell types and early regeneration in SKP seeded grafts was comparable to those seeded with SCs. Histomorphometrical and electrophysiological measurements of cell-treated nerve segments after 8 weeks survival all showed significant improvement as compared to diluent controls. A possible mechanistic explanation for the observed results of improved regenerative outcomes lies in SKP–SCs' ability to secrete bioactive neurotrophins. We therefore conclude that SKPs represent an easily accessible, autologous source of stem cells for transplantation therapies which act as functional Schwann cells and show great promise in improving regeneration following nerve injury.

Section snippets

Cell culture

SKPs were generated from dermis of postnatal day 2 Lewis rats and cultured according to published protocols (Toma et al., 2001). Briefly, pups were quickly decapitated and skin on the dorsal torso was sterilized with a 70% EtOH swab prior to removal with sterile scissors. Collected tissue was minced in Hank's balanced salt solution (HBSS; GIBCO, Burlington, ON, Canada) on ice and then incubated for approximately 45 min in 0.1% collagenase at 37 °C. Skin pieces were mechanically dissociated,

Results

In all animals, the surgical procedures were well-tolerated and wounds healed without complication or signs of pain or discomfort. Graft repair remained intact in every case and there was no evidence of excessive fibrosis in any of the groups.

Discussion

The major findings in this study were (1) Skin-derived precursor cells pre-differentiated towards a SC phenotype in vitro survived, migrated, and maintained the expression of SC markers for at least 8 weeks when transplanted into an acellular (freeze-thawed) isografts (2) After 4 weeks SKP–SCs supported axonal elongation to the same extent as autograft and SC treatment; and (3) provided superior axon regeneration, myelination, and electrophysiological recovery as compared to media control at

Acknowledgments

The authors thank Shahbaz Syed, Joanne Forden and Dr. Qing Gui Xu for their technical expertise and assistance with the preparation of this manuscript. We thank Dr. Freda Miller and her laboratory personnel for demonstrating the techniques of SKP culture and generation. This research was supported by a grant from the Canadian Institutes for Health Research (MOP 82726) and an NSERC CGSD studentship granted to S.K.W. The authors have no conflict of interest.

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