Biocompatibility Of Functionalized Self-assembling Peptide Hydrogel With Neural Stem Cells

Objective:To culture rat neural stem cells (NSCs) the in vitro. Methods:NSCs harvested from the cerebral cortex of neonatal one-day rats were triturated and cultivated in serum-free medium. The morphology of growth of cells was demonstrated by inverted phase contrast microscopy. Immunofluorescence staining with Nestin was used to identify NSCs. Result:The cells isolated and cultured grew in a manner of neurospheres and suspended in the medium. The cells were capable of proliferating and maintaining long-term survival in vitro. Conclusion:NSCs derived from cerebral cortex of neonatal rats were successfully isolated and cultured in vitro.Objective:To design and synthesize a new self-assembling peptide hydrogel scaffold.Methods:Peptide RADA16 and RADA16-IKVAV were synthesized by solid phase method. The functionalized self-assembling peptide named IKVAVmx was then made by mixing the above two peptide solutions. The morphological features of IKVAVmx were studied by atom force microscope (AFM). Result:IKVAVmx could self-assemble to form a hydrogel. AFM showed that self-assembly hydrogel was made up of interconnected nanofibers.Conclusion:IKVAVmx peptide was synthesized and could self-assemble into hydrogel.Objective:To observe the of functionalized self-assembling peptide hydrogel with neural stem cells in vitro. Methods:NSCs were isolated from the cerebral cortex of neonatal rats and cultured in serum-free medium. NSCs were seeded on the surface of both the functionalized self-assembling peptide hydrogel (experimental group) and RADA16 self-assembling peptide hydrogel (control group). The proliferation behavior of the cells was evaluated by CCK-8 assay. The cell migration was detected by laser scan confocal microscope. Immunofluorescence staining with Nestin, MAP2, GFAP, and CC-1 was used to assess the differentiation of NSCs. The protein expression of MAP2 was determined by Western blot analysis. Result:The power of cell proliferation and migration in experimental group was significantly higher than that of control group (P<0.05). Compared with control group, the percentage of MAP2 positive cells increased significantly (P<0.05) and the percentage of GFAP positive cells decreased significantly (P<0.05) in experimental group. Western blot analysis indicated that MAP2 activity in IKVAVmx scaffold was significantly higher than that in pure RADA16 scaffold (P<0.05). Conclusion:The functionalized self-assembling peptide hydrogel has good biocompatibility with NSCs, suggesting that it may be a promising biomaterial for neural tissue engineering.