The Development of a Cryopreservation Method Suitable for a Large Cyanobacteria Collection

Laboratories worldwide working with cyanobacteria have created culture collections to carry out related studies. However, the lack of manpower (especially after the studies were finished), maintenance costs and proper preservation methods often results in the loss of research materials and a waste of isolation effort. Several parameters are generally considered very important in cryopreservation, including the choice of the cryoprotectant, cryoprotectant concentration, freezing rate, physiological status of the culture and thawing procedure. This makes it very difficult to establish universal guidelines for cyanobacteria cryopreservation. Herein, we present a cryopreservation method suitable for a range of strains, using two cryoprotectants (methanol and dimethyl sulfoxide at a final concentration of 5 and 3 %, respectively) along with a combined vital staining and reproductive viability criteria for the post-thawing recovery. The obtained results are very encouraging as more than 83 % of tested cyanobacteria were amenable for cryopreservation, 80 % of strains (111 in total) showing more than 90 % recovery with at least one of the cryoprotectants used.