This study presents the influence of control parameters including population (NP) size, mutation factor (F), crossover (Cr), and four types of differential evolution (DE) algorithms including random, best, local-to-best, and local-to-best with self-adaptive (SA) modification for the purpose of optimizing the compositions of dimethylsufloxide (DMSO)-free cryoprotectants. Post-thaw recovery of Jurkat cells cryopreserved with two DMSO-free cryoprotectants at a cooling rate of 1 °C/min displayed a nonlinear, four-dimensional structure with multiple saddle nodes, which was a suitable training model to tune the control parameters and select the most appropriate type of differential evolution algorithm. Self-adaptive modification presented better performance in terms of optimization accuracy and sensitivity of mutation factor and crossover among the four different types of algorithms tested. Specifically, the classical type of differential evolution algorithm exhibited a wide acceptance to mutation factor and crossover. The optimization performance is more sensitive to mutation than crossover and the optimization accuracy is proportional to the population size. Increasing population size also reduces the sensitivity of the algorithm to the value of the mutation factor and crossover. The analysis of optimization accuracy and convergence speed suggests larger population size with F > 0.7 and Cr > 0.3 are well suited for use with cryopreservation optimization purposes. The tuned differential evolution algorithm is validated through finding global maximums of other two DMSO-free cryoprotectant formulation datasets. The results of these studies can be used to help more efficiently determine the optimal composition of multicomponent DMSO-free cryoprotectants in the future.