In this study, we investigated the effect of solid culture medium on the production of cordycepin in Cordyceps militaris. The regression equation was expressed as follows: Y1= 755.3-58.6625X1+4.79432E-14X2-46.6625X3-5.66269E-14X1X2- 0.025X1X3+1.62475E-14X2X3-160.6625X1 2+0.0125X2 2-206.9625X3 2, where, Y represents the value of cordycepin content (μg/g), X1 corresponds to the weight of M. alternatus in solid culture medium (g/bottle), X2 to the water content of the solid culture medium (%), and X3 to the culture period (day). The solid culture medium was optimized using the response surface methodology, and the optimal medium composition was as follows: the weight of M. alternatus in solid culture medium and water content were 16.2% and 100.7% (20.14 mL water/20 g solid culture medium), respectively, with a culture period of 39 days. Under these conditions, the cordycepin content of the fruiting bodies reached 150.0 μg/g (actual value). The supplementation of M. alternatus in solid culture for improved cordycepin content of C. militaris seems to be a promising alternative to wild and solid cultivation.