In order to promote the industrialization of Lactobacillus plantarum BF_15 with multiple probiotic properties (such as immune regulation and cholesterol reduction) from breast-fed infants with clear genetic background. In this study, the medium components and culture conditions were optimized through single-factor combined with orthogonal experiments, and the lyoprotectants were optimized through single-factor experiments combined with specific formulation ratios. The results showed that the viable cell count of BF_15 in MRS Medium was 2.30×109 CFU/mL. The optimal medium composition obtained through singlefactor combined orthogonal experiments was: fructooligosaccharides, 30 g/L; nitrogen source (soy protein peptone: yeast extract=3:1), 30 g/L; sodium acetate anhydrous, 6 g/L; manganese sulfate, 75 mg/L. Under these conditions, the viable cell count of BF_15 reached 6.93×109 CFU/mL. The optimal culture conditions: inoculation volume fraction, 5%; incubation temperature, 33 ℃; initial pH value 7.0; dissolved oxygen level, 0 r/min. Under such conditions, a viable cell count of 8.92×109 CFU/mL was achieved, which was 3.88 times that before optimization. The optimal lyoprotectant formulation was: skim milk, 100 g/L; bovine serum, 10 ml/L; sucrose, 80 g/L; sodium glutamate, 20 g/L; yeast extract powder, 15 g/L; Tween 80, 10 ml/L. Under these conditions, the survival rate of BF_15 was 90.57%, with a viable cell count of 2.45×1010 CFU/mL. After a storage at 4 ℃ for 56 days, the survival rate of the freeze-dried bacterial powder was 82.35%, with a viable cell count of 2.02×1010 CFU/mL. Following the gastrointestinal fluid treatment, the survival rate was 69.70%, with a viable cell count of 1.41×1010 CFU/mL. In summary, the high-density culture of Lactobacillus plantarum BF_15 was realized and highly viable bacteria preparations were obtained, which provides solid theoretical support for accelerating the industrialization process.