The invention relates to a method for constructing a three-dimensional
neural stem cell model in two steps by adopting the micro-fluidic technology. The method is characterized in that a
rat tail collagen I is used as a three-dimensional support, a micro-pillar array type micro-fluidic
chip is used as a culture platform, and a
neural stem cell is cultured in two steps, wherein in the early culture stage, a culture medium for promoting the amplification of the
neural stem cell is injected into a
cell culture chamber, and in the later culture stage, a
conditioned medium suitable for the growth of the neural
stem cell and the
daughter cells thereof is used, and a three-dimensional
composite structure which is similar to a nerve tissue is formed by simulating the microenvironment of different
neurogenesis stages in the body. The method provided by the invention is good in
repeatability and can be used for construction a plurality of groups of samples. The adopted microfluidic culture
system is in a microliter volume and can be regulated accurately, thus the amount of various high-cost
cell growth factors, immunologic fluorescent antibodies and
cell hormones used in the process of culturing the cell can be reduced greatly, and the
cell culture cost can be lowered. The three-dimensional neural
stem cell model is expected to be a nerve tissue substitute for screening a novel medicament or monitoring an environmental
toxin.