Volume 21, Number 110 (5-2014)                   daneshvarmed 2014, 21(110): 69-76 | Back to browse issues page


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Transfection of BMSCs by pSec-TAG-A-CNTF for spinal cord injury. daneshvarmed. 2014; 21 (110) :69-76
URL: http://daneshvarmed.shahed.ac.ir/article-1-884-en.html

Abstract:   (3266 Views)
Background and Objective: Ciliary neurotrophic factor (CNTF) was originally described as a trophic factor supporting the survival of ciliary ganglion neurons in vitro. CNTF can also promote the survival and differentiation of a variety of nerve cells including sensory, motor, autonomic neurons and oligodendrocytes. To enhance the neurotrophic effect of BMSCs, the CNTF-pSec-tag 2 /hygro A plasmid was constructed and transferred into BMSCs by lipofectamin. Materials and Methods: Plasmid CNTF-pSec-tag 2 /hygro A was constructed and verified by sequencing. BMSCs were transfected by lipofectamin. The transcription of the gene was evaluated by RT-PCR and real time PCR, and the expression of protein was evaluated by western blotting and ELISA. Results: CNTF-pSec-tag 2 /hygro A plasmid was correctly verified. After transfection, the transcription of CNTF gene and the expression of CNTF protein were proven by RT-PCR, western blotting and ELISA. Conclusion: CNTF-pSec-tag 2 /hygro A plasmid was correctly constructed and BMSCs were successfully transfected by transfection and CNTF-pSec-tag 2 /hygro A protein can be expressed well, which is a good foundation for future studies on the transplantation of gene-modified BMSCs to promote spinal cord regeneration
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Type of Study: Research | Subject: فیزیولوژی ورزش

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