Growth and differentiation of neural stem cells in a three-dimensional collagen gel scaffold

摘要

Collagen protein is an ideal scaffold material for the transplantation of neural stem cells. In this study, rat neural stem cells were seeded into a three-dimensional collagen gel scaffold, with suspension cultured neural stem cells being used as a control group. Neural stem cells, which were cultured in medium containing epidermal growth factor and basic fibroblast growth factor, actively expanded and formed neurospheres in both culture groups. In serum-free medium conditions, the processes extended from neurospheres in the collagen gel group were much longer than those in the suspension culture group. Immunofluorescence staining showed that neurospheres cultured in collagen gels were stained positive for nestin and differentiated cells were stained positive for the neuronal marker βIII-tubulin, the astrocytic marker glial fibrillary acidic protein and the oligodendrocytic marker 2′,3′-cyclic nucleotide 3′-phosphodiesterase. Compared with neurospheres cultured in suspension, the differentiation potential of neural stem cells cultured in collagen gels increased, with the formation of neurons at an early stage. Our results show that the three-dimensional collagen gel culture system is superior to suspension culture in the proliferation, differentiation and process outgrowth of neural stem cells. Research Highlights (1) The effect of collagen gels on the process outgrowth of neural stem cells was observed in this study. The results confirmed that neural stem cells could be passaged in a three-dimensional collagen gel scaffold. (2) After culture for 4, 7, and 14 days, the neural stem cells cultured in a three-dimensional collagen gel or in suspension were selected for induced differentiation experiments, which showed that three-dimensional collagen gel scaffolds can promote the differentiation of neural stem cells into neurons.