Background: Platelet-rich plasma (PRP) has been widely used to improve wound healing and tissue repair.The objective of this study was to assess a cycle of freezing/thawing and its effect in the amount of growth factors in PRP.Materials and methods: After the preparation of PRP using standard methods, the samples were equally divided into two groups: The first group was activated by adding 10% calcium gluconate, while the second group was cryopreserved at -80°C for 24 hours.Enzyme-linked immunosorbent assay was used for aliquots of PRP to measure concentrations of insulin-like growth factor-1, platelet-derived growth factor, and basic fibroblast growth factor.Results: 15 mL of whole blood was obtained from 20 volunteers and collected into citrated tubes.The mean platelet count of the donors and the autologous PRP were 238.5 ± 44.7 × 10 3 /μL and 544.7 ± 161.5 × 10 3 /μL, respectively.There were no significant differences between the growth factor levels of freeze-thawed and calcium-activated PRP.A cycle of freezing/thawing was the only independent factor associated with the growth factor yield in the multivariate model. Conclusion:With its features of being simple, inexpensive, and easy for standardization, a cycle of freezing/thawing may be the method of choice for PRP activation procedure, without inducing fibrin matrix.This study was performed in a laboratory setting; therefore, future clinical trials are recommended.