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. 2008 May 22;24(2):57. doi: 10.1007/s12264-008-0057-y

Ex vivo non-viral vector-mediated neurotrophin-3 gene transfer to olfactory ensheathing glia: effects on axonal regeneration and functional recovery after implantation in rats with spinal cord injury

神经营养素 — 3 基因非病毒载体转染的嗅鞘细胞移植促进脊髓损伤大鼠轴突再生及功能恢复

Jun Wu 1, Tian-Sheng Sun 1,✉, Ji-Xin Ren 1, Xian-Zhang Wang 1
PMCID: PMC5552512  PMID: 18369383

Abstract

Objective

Combine olfactory ensheathing glia (OEG) implantation with ex vivo non-viral vector-based neurotrophin-3 (NT-3) gene therapy in attempting to enhance regeneration after thoracic spinal cord injury (SCI).

Methods

Primary OEG were transfected with cationic liposome-mediated recombinant plasmid pcDNA3.1(+)-NT3 and subsequently implanted into adult Wistar rats directly after the thoracic spinal cord (T9) contusion by the New York University impactor. The animals in 3 different groups received 4×105 OEG transfected with pcDNA3.1(+)-NT3 or pcDNA3.1(+) plasmids, or the OEGs without any plasmid transfection, respectively; the fourth group was untreated group, in which no OEG was implanted.

Results

NT-3 production was seen increased both ex vivo and in vivo in pcDNA3.1(+)-NT3 transfected OEGs. Three months after implantation of NT-3-transfected OEGs, behavioral analysis revealed that the hindlimb function of SCI rats was improved. All spinal cords were filled with regenerated neurofilament-positive axons. Retrograde tracing revealed enhanced regenerative axonal sprouting.

Conclusion

Non-viral vector-mediated genetic engineering of OEG was safe and more effective in producing NT-3 and promoting axonal outgrowth followed by enhancing SCI recovery in rats.

Keywords: functional recovery, gene therapy, neurotrophin-3, olfactory ensheathing glia, regeneration, spinal cord injury, non-viral vectors

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