A Chinese hamster ovary (CHO) cell line is the primary choice for the production of recombinant protein drugs with glycosylation modification. Recombinant protein productivity was enhanced by the addition of dimethyl sulfoxide (DMSO) in CHO cells. However, DMSO induced G0/G1 phase growth arrest and reduced cell growth rate. A two-stage process was developed to utilize the stimulatory effect of DMSO on recombinant protein production and mitigate the problem of growth inhibition in this study. In the first stage, cells are cultured without DMSO for a certain period in order to obtain a high cell density. Sequentially, DMSO is added to achieve a high specific productivity in the second stage. Using this approach, we found that by adding 1% DMSO after 24 h of growth, macrophage colony-stimulating factor (M-CSF) volumetric productivity is increased by 57% compared with the value obtained without the addition of DMSO.