The medium for artificial culture is very important for the survival and development of biocrusts. The physiological responses of biocrusts to the addition of saccharides and growth regulators were investigated to evaluate the optimal cultivation method. The research results suggested that MDA was the lowest and chlorophyll-a, soluble protein, and soluble sugar were the highest under the addition of 0.5 mg/L sucrose and glucose (10 mg/L). The MDA content of the biocrusts decreased with increasing 2,4-D and 6-BA concentrations. Chlorophyll-a, soluble protein, and soluble sugar of the biocrusts were the most abundant when the additions of 2,4-D and 6-BA were both 0.5 mg/L. The chlorophyll-a concentration of the biocrusts always increased with the addition of sucrose and glucose. The maximum tolerance of biocrust to sucrose was 40 mg/L. The excessive sugar in the medium had an inhibitory effect on the growth and reproductive cycles and resulted in greater damage to moss cells in biocrusts as sugar concentrations increased. The appropriate addition of saccharides and growth regulators can promote the survival and growth of desert biocrusts.