A new rolling culture system with double-mouthed roller was manufactured. Hybridoma, which produces anti-hCG monoclonal antibody (mAb), was employed in the double-mouthed roller, in which growth and antibody yield of the cells were compared with those cultivated in T flask. The results showed that with an increase in growth of the cells cultured in the roller by 1.4～1.8 folds, the yield of the antibody increased by 1.3 folds of those statically cultured in T flask. Addition of gelatin in culture media could further improve both yield of the antibody and growth of the hybridoma cells.