The invention relates to a preparation method of photoresponsive intelligent gel microspheres for three-dimensional
cell culture. The preparation method is characterized by comprising the following steps: dissolving poly(
methyl vinyl ether-
maleic anhydride) and beta-
cyclodextrin into a
solvent, namely N,N-
dimethylformamide, carrying out heating reaction under the protection of
nitrogen, and removing the
solvent; adding a mixture water solution of
polyethylene glycol 20000 and
agarose into the reactant, and vibrating until the reactant is dissolved, so as to prepare a water-phase solution for later use; dropwise adding the water-phase solution into oil-phase
simethicone, carrying out heating, stirring, reaction and standing, removing supernate, cleaning microspheres deposited at the bottom of a container, and carrying out
vacuum drying, so as to obtain the gel microspheres; putting the gel microspheres into Azo-RGD
methane water solution, vibrating for 24 hours under the
room temperature, repeatedly rinsing by virtue of deionized water, and
drying by virtue of N2. By utilizing the gel microspheres prepared by virtue of the preparation method, growing environments of cells
in vivo can be simulated, and the adsorption and controllable separation of the cells on the microspheres can be realized.