Abstract：A simple and optimized cryopreservation process was applied to Cistanche deserticola callus. To obtain optimal results, the callus was pre-cultured on B5 medium supplemented 6% (j) dimethyl sulfoxide (DMSO) and then treated with a vitrification solution at 25℃ for 20 min before immersion in liquid nitrogen. The vitrification solution contained 30% (j) glycerol, 15% (j) ethylene glycol, 10% (j) DMSO and 0.5 mol/L sucrose. After cryopreservation and rapid thawing in a water bath at 30℃, the callus was washed with 1.0 mol/L sucrose solution at 25℃. The survival rate of callus was 86% under the above cryopreservation process. After the cryopreserved callus was cultured on B5 medium for five months, the content and production of PeG were 97% and 95% of those in the non-frozen callus.