Basic & Clinical Medicine ›› 2022, Vol. 42 ›› Issue (1): 75-81.

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PF-127-LV-NTFs three-dimensional composite scaffold in culturing rat neural stem cells

  

  • Received:2021-09-27 Revised:2021-11-19 Online:2022-01-05 Published:2022-01-05
  • Contact: Hong-Fu WU E-mail:hongfuw@126.com

Abstract: Objective To investigate the effect of LINGO-1 shRNA (LV) and neurotrophic factors cocktail (NF,NFcocktail) encapsulated by Pluronic F-127(PF-127)hydrogel three-dimensional composite scaffolds on proliferation and differentiation of rat neural stem cells. Methods The LV was constructed and infected with NSCs at different MOI. RT-qPCR and Western blot were used to detect the expression of LINGO-1 in NSCs. BDNF NT-3 PDGF IGF-1 EGF bFGF and GDNF groups were divided into NFcocktail. The LV and NFcocktail were mixed with PF-127 and divided into different groups to culture NSCs. CCK-8 was used to detect the of cell viability of NSCs, the survival rate of NSCs was detected by LIVE/DEAD staining, and immunofluorescence staining was used to detect the proliferation and differentiation of NSCs. Results the optimal MOI for LV was 5. CCK-8 assay results showed that there was no significant difference in the survival rate of NSCs. The LIVE/DEAD and Nestin immune-fluorescence staining results showed that compared with control, the survival rate of NSCs and the proportion of neuron differentiation positive were the highest in PF-127+LV+NF group(P<0.01). Conclusions PF-127-LV-NFcocktail three-dimensional composite scaffold is beneficial to the proliferation and the directional differentiation of NSCs in rat.

Key words: neural stem cells, LINGO-1 shRNAl, PF-127 hydrogel, neuronal differentiation

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