摘要
Objective:To construct a recombinant lentiviral vector that co-express ZsGreen and BDNF and establish an Adipose derived stem cell (ADSC) line with stable BDNF expression down- regulation.Methods: BDNF were designed and inserted into the lentiviral vector p HBLV-CMVIE-Zs Green-Puro. After identification by DNA sequencing, the lentiviral vectors carrying BDNF were packaged in 293 cells. The transfection efficiency was observed under fluorescence microscope;the lentiviral particles were collected to infect mouse ADSCs, and BDNF level were assessed using real-time PCR and Western blotting.Results: DNA sequencing demonstrated successful construction of the BDNF lentivirus vectors. Real- time PCR and Western blotting showed a high level BDNF mRNA and protein.Conclusion: We have successfully established an ADSC model with stable BDNF expression which establish the basis for the potential application of ADSCs-BDNF in the treatment of AD.
基金
The National Natural Science Foundation of China.Project No:(81660224,81100246)
Hainan Provincial Key Research and Development Project.Project No:(ZDYF2017120,ZDYF2016110)
National College’s Students’Innovation and Entrepreneurship Training Program Project.Project No:(201711810010,HYCX2016010,HYCX2018032)
Hainan Provincial Health and Family Planning Industry Research Project.Project No:(18A200115)
Haikou People’s Hospital Project.Project No:(2018-SNP-01-003).
