Abstract: By using precipitation polymerization，thermo-sensitive poly(N-isopropylacrylamide-co- glycidyl methacrylate) [P(NIPAM-co-GMA)] microspheres were synthesized via the one-step and two-step reaction methods. The one-step method proceeded with the polymerization reaction of all reactants at the same time，while the two-step method proceeded by the polymerization reaction of NIPAM firstly，and then adding GMA to form P(NIPAM-co-GMA) microspheres. The poly(N- isopropylacrylamide-co-2- hydroxypropyl methacrylate/ethylenediamino-β-cyclodextrin) [P(NIPAM-co- GMA/β-CD)] microspheres were fabricated through chemical reaction of P(NIPAM-co-GMA) microspheres with modified ethylenediamino-β- cyclodextrin (EDA-β-CD). The morphologies，chemical structures and thermo-sensitive properties of all prepared microspheres were characterized by scanning electron microscopy，Fourier transform infrared spectroscopy，dynamic light scattering and microscope. The results showed that the prepared microspheres by both methods had good monodispersity and sphericity. The microspheres with β-CD groups were prepared successfully，and all microspheres had good thermo-sensitive properties. In addition，microspheres prepared by the two-step method were larger and hadve more pendent β-CD groups than that prepared by the one-step method.