Combined Application of Neutrophin-3 Gene and Neural Stem Cells is Ameliorative to Delay of Denervated Skeletal Muscular Atrophy after Tibial Nerve Transection in Rats

Cell Transplantation

Article

Yang Wang¹  Lei Xu²  Shaonan Hu²  Jianguang Xu²  Yudong Gu²  Changqing Zhang³  Sen Lin⁴ 

[1] Department of Anatomy, Histology and Embryology, Shanghai Medical Center, Fudan University, Shanghai, China;Department of Hand Surgery, Huashan Hospital, Fudan University, Shanghai, China;Department of Orthopaedics, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, China;Department of Orthopaedics, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, China; Department of Hand Surgery, Huashan Hospital, Fudan University, Shanghai, China;

关键词: Neural stem cells (NSCs);    Neurotrophin-3 (NT-3);    Denervated skeletal muscular atrophy;    Transplantation;   

DOI  :  10.3727/096368910X524773

 received in 2010-04-10, accepted in 2010-07-23,  发布年份 2011

来源: Sage Journals

【 摘 要 】

Examination of the therapeutic efficacy of neural stem cells (NSCs) has recently become the focus of much investigation. In this study we present an insight of the effects of combined application with neurotrophin-3 (NT-3) and NSCs that derived from rat embryo spinal cord on delaying denervated skeletal muscular atrophy after tibial nerve was severed. NT-3 gene was amplified by PCR and subcloned into lentiviral vector pWPXL-MOD to construct a lentiviral expression vector pWPXL-MOD-NT-3. A positive clone expressing NT-3 (named NSCs-NT-3) was obtained and used for differentiation in vitro and transplantation. Sixty adult rats, whose tibial nerves were sectioned, were divided into two groups: one grafted with NSCs-NT-3 (experimental group, n = 30) and the other with NSCs transfected by pWPXL-MOD (control group, n = 30). The cell survival and differentiation, NT-3 gene expression, and effect of delaying denervated skeletal muscular atrophy were examined through immunohistostaining, RT-PCR, Western blot, electrophysiological analysis, and mean cross-sectional area (CSA) of gastrocnemius, respectively. The results show that the NT-3 gene, which is comprised of 777 bp, was cloned and significantly different expression were detected between NSCs and NSCs-NT-3 in vitro. Quantitative analysis of the choline acetyltransferase (ChAT) immunopositive cells revealed a significant increase in experimental group compared to the control group 4 weeks after implantation (p < 0.01). Twelve weeks after transplantation, the ChAT immunopositive cells were detected near the engrafted region only in experimental group. Furthermore, the effect in delaying denervated skeletal muscular atrophy is indicated in the EMG examination and mean CSA of gastrocnemius. These findings suggest that the neural stem cells expressing NT-3 endogenously would be a better graft candidate for the delay of denervated skeletal muscular atrophy.

【 授权许可】

Unknown   
© 2011 Cognizant Comm. Corp.

【 图 表 】

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Figure 7.