Nanoporous cellulose films was prepared using multiple freeze/thaw cycles method and subsequent surface hydrophobicity modification by coconut oil. Firstly, HCl depolymerized cotton fiber waste was dissolved in NaOH/urea mixture solvent. Then, subzero-temperature cellulose solution was cast onto acrylic mold and thawed to room temperature to obtain cellulose gel. Solvent exchange with water was repeatedly carried out to obtain cellulose gel film. The resultant cellulose gel film was treated with a concentration of glycerol to prevent the film brittleness. Finally, glycerol treated cellulose film was subject to multiple freeze/thaw cycles to obtain nanoporous cellulose film and then the modification was carried out by coconut oil. Nanoporous structure was identified by SEM and surface hydrophobicity were determined using ATR-FTIR spectroscopy which revealed the strong fatty ester peak at 1744 cm-1. In summary, cellulose films having the nanoporous structure was successfully prepared using multiple freeze/thaw cycles method. It was found that dimension stability (shrinkage), pore sizes of nanoporous cellulose films and cholesterol adsorption were dependent on concentrations of cellulose solution, concentrations of glycerol and a number of freeze/thaw cycles. The results indicated that the optimal adsorption was found with the nanoporous cellulose film prepared from 7 wt% cellulose solution, 30 wt% glycerol solution, and 5 freeze/thaw cycles.