Abstract: Objective To compare the three different methods of dispersing neuropheres of neural stem cells into single cells.Methods The neural stem cells were isolated of 16～20 weeks old human fetus,and were cultured and expanded to form neuropheres.Three different dispersing methods including digesting with Trypsin,triturating with Pasteur pipette and the new method of digesting and shaking were used to disperse neuropheres and the viability and clonability of the resulting cells were analyzed.Results The method of digesting combined with shaking yielded the best survival rate and best proliferation potential of cells.Conclusion This study suggests a valid and safe method that can be used for efficient passage of neural stem cells.

Key words: human, embryo, neural stem cells, neuropheres