Combined treatment of neurotrophin-3 gene and neural stem cells is propitious to functional recovery after spinal cord injury.
We present an insight of the effects of combination therapy with neurotrophin-3 and neural stem cell on functional recovery after spinal cord injury (SCI). Total RNA was extracted from neural stem cell line C17.2 and reversed transcribed into cDNA. Neurotrophin-3 (NT-3) gene was amplified by PCR and subcloned into plasmid to construct an expression vector pNT-3. A positive clone containing pNT-3, named SHN2, was obtained and used for transplantation. Thirty adult mice received mechanical injury at the T8 vertebra level. Cell survival, NT-3 gene expression, and functional recovery were observed through X-Gal staining, RT-PCR, and open field locomotion, respectively. The results show that NT-3 gene comprising 777 bp nucleotides was cloned and a more than twofold expression was detected when transfected into neural stem cell line C17.2. Quantitative analysis of cellular density revealed a significant increase in SHN2 compared to the control cells (p < 0.01). Thirty days after transplantation, SHN2 showed significant increase near the lesion site. Furthermore, the functional recovery indicated an active effect by detecting Basso-Beattie-Bresnahan (BBB) locomotor rating scale (p < 0.01). In conclusion, combined treatment of neural stem cells and NT-3 gene can facilitate functional recovery. It offers an effective approach to treat SCI.
Zhang
Luyi
L
Laboratory of Molecular Neural Biology, School of Life Sciences, Shanghai University, Shanghai 200444, China.
Gu
Shuting
S
Zhao
Cuiping
C
Wen
Tieqiao
T
D000818:Animals; D001522:Behavior, Animal; D002460:Cell Line; D049109:Cell Proliferation; D003001:Cloning, Molecular; D003131:Combined Modality Therapy; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D051379:Mice; D016501:Neurites / Q000378:metabolism; D009474:Neurons / Q000637:transplantation*; D020933:Neurotrophin 3 / Q000235:genetics*; D020127:Recovery of Function; D013119:Spinal Cord Injuries / Q000235:genetics* / Q000628:therapy*; D033581:Stem Cell Transplantation
D020933:Neurotrophin 3