Magnetic hybrid hollow spheres of about 200 nm were prepared by a core-template-free route, that is, adding Fe3O4 nanoparticles stabilized by poly(vinyl alcohol) (PVA) to an aqueous solution of polymer-monomer pairs composed of a cationic polymer, chitosan (CS), and an anionic monomer, acrylic acid (AA), followed by polymerization of acrylic acid and selective cross-linking of chitosan at the end of polymerization. The obtained hybrid spheres were characterized by dynamic light scattering (DLS) in aqueous solution and observed by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and atomic force microscopy (AFM) in the solid state. Fourier transform infrared spectroscopy (FTIR) and X-ray and electron diffractions revealed that the Fe3O4 nanoparticles were incorporated into the shells of chitosan- poly(acrylic acid) (CS-AA) hollow spheres. Magnetization studies and Mossbauer spectroscopy suggested that the chains (or islands) of iron oxide nanoparticles were most likely formed in the walls of the hollow spheres. The phantom test of magnetic resonance imaging showed that the synthesized hybrid hollow spheres had a significant magnetic resonance signal enhancement in T2-weighted image.