A series of cellulose films with rough surface were prepared by employing a simple solution casting method and using the waterproof abrasive papers with different grits as the substrate. The cellulose films possessed a rough surface with the maximum height difference (Sz) of 128-217 μm, a macroporous structure with a high porosity of 84.7%-90.5%, and a negative potential between -40.00 and -54.15 mV. Furthermore, the cellulose films exhibited excellent microalgae adhesion properties. After 18 days of attached Chlorella sp. cultivation experiments, the average productivities of C-A-120 films (C-A-X, X means the mesh number of the substrate) reached 20.80 g m-2 d-1), which is 2.69 times than that of the cellulose film with a smooth surface. The result indicates that the cellulose films with a rough surface and high water adsorption ratio have a huge potential in serving as the substrate of the attached microalgae cultivation to promote microalgae cells growth and biofilm formation.