Cryopreservation of the Microalgae Scenedesmus sp.

Abstract

Each phytoplankton species presents a different behavior and tolerance to the cryopreservation process. Therefore, in a species-specific protocol, it is essential to ensure both growth and post-thawing cell viability. In this study, we explored the effect of cryopreservation of Scenedesmus sp. with two cryoprotectants, dimethyl sulfoxide (DMSO) and methanol (MET), at 5% and 10% inclusion for each. In the control treatment, the microalgae were not exposed to cryoprotective agents (Control). Three post-thawing cell viability criteria were used: no cell damage (NCD), cell damage (CD), and marked lesions (LM), and mitochondrial and cell membrane damage was evaluated by flow cytometry. The study was a 2 × 2 factorial design, with five replications by treatments, population growth, and cell damage evaluated from the fifth day after thawing. On the fifth day, the highest percentage of NCD was observed when the microalgae were cryopreserved with DMSO 5% (50%); Regarding the control group, it showed 0% NCD. Flow cytometry analysis reveals minor damage at the membrane and mitochondria (9–10.7%) when DMSO is used at both inclusion percentages (5–10%) after thawing. In the exponential phase, the highest growth rates, doubling time, and yield was observed in cryopreserved cells with MET 5%. The results suggest that DMSO 5% is an ideal treatment for cryopreserving microalgae Scenedesmus sp.