تراسفکشن پایدار سلول‌های استرومایی مغز استخوان توسط وکتورpSecTag2/HygroA-CNTF به‌منظور استفاده در درمان ضایعات نخاعی

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