Whey based medium was optimized for the production of viable Lactobacillus crispatus KLB 46 isolated from the vagina of Korean women. Among the various nitrogen sources such as yeast extract, beef extract, and proteose peptone no. 3 supplemented to whey, beef extract showed the highest viable cell production. The addition of Tween 80 to the whey based medium increased viable cell concentration. As beef extract supplementation is not economically attractive, corn steep liquor was added as a supplementary nitrogen sources. When corn steep liquor was supplied with beef extract with the ratio 5 : 1, the viable cell count was $3.11{ imes}10^9$ CFU/mL. Also, the addition of mineral salts containing sodium acetate (5 g/L), potassium phosphate dibasic (2 g/L), magnesium sulfate (0.1 g/L) and manganese sulfate (0.05 g/L) to the whey medium increased viable cell count further ($5.00{ imes}10^9$ CFU/mL).