Abstract
BackgroundThe cell source for transplantation therapy is always a prerequisite question to be solved in clinical applications. Neural cells are considered non-regenerable, which highly restrict their application in the treatment for nerve injury. Therefore, neural trans-differentiation based on gene transfection provides a new solution to this issue. Compared to viral strategy, non-viral gene delivery systems are considered as a more promising way to achieve this aim. This study centers on a novel application of Porphyra yezoensis polysaccharide as a non-viral gene carrier for the neural trans-differentiation of mouse fibroblasts.ResultsEthanediamine modified P. yezoensis polysaccharide (Ed-PYP) served as a gene carrier and a group of plasmids that encode Ascl1, Brn4, and Tcf3 (pABT) self-assembled into nanoparticles. Results demonstrated that Ed-PYP–pABT nanoparticles at Ed-PYP: pABT weight ratio of 40:1 was the optimal candidate for gene delivery. ELISA assay revealed the highest expression levels of NGF, BDNF and SHH at 14 days after last transfection. Immunofluorescence and western blot assays also showed robust expression of neural markers including Nestin, GFAP, β-3tubulin, NF200, GAP43 and MAP2, in induced 3T6 cells at this time point.ConclusionOverall, these findings indicated that the P. yezoensis polysaccharide-based non-viral gene co-delivery system is a promising strategy for the generation of neural cells, which might facilitate the developments in the recovery of neural injuries.
Highlights
The cell source for transplantation therapy is always a prerequisite question to be solved in clinical applications
The ethylenediamine modified Porphyra yezoensis polysaccharide (Ed-Porphyra yezoensis polysaccharide (PYP))– pABT was prepared at different MEd-PYP/MpABT weight ratios while the group of free plasmids Ascl1, Brn4 and Tcf3 was applied as negative control in this study
A higher Ed-PYP: pABT weight ratio resulted in a better combination of Ed-PYP and pABT
Summary
The cell source for transplantation therapy is always a prerequisite question to be solved in clinical applications. Non-viral gene delivery systems are considered as a more promising way to achieve this aim. This study centers on a novel application of Porphyra yezoensis polysaccharide as a non-viral gene carrier for the neural trans-differentiation of mouse fibroblasts. Reprogramming of somatic cell lines is considered as a promising method for cell therapy and regenerative medicine. Viral gene vectors, possessing high transfection efficiency, have been widely used for gene delivery; the clinical application of viral vectors is hampered by serious drawbacks such as abnormal gene expression [8,9,10], immune responses [11,12,13], limitations with respect to scale-up procedures, and inflammation [14]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
