title: Comparative analysis of cryopreservation methods in Chlamydomonas reinhardtii.
abstract: Chlamydomonas is a model organism used for studies of many important biological processes. Traditionally, strains have been propagated on solid agar, which requires routine passaging for long-term maintenance. Cryopreservation of Chlamydomonas is possible, yet long-term viability is highly variable. Thus, improved cryopreservation methods for Chlamydomonas are an important requirement for sustained study of genetically defined strains. Here, we tested a commercial cryopreservation kit and directly compared it's effectiveness to a methanol-based method. We also tested thaw-back procedures comparing the growth of cells in liquid culture or on solid agar media. We demonstrated that methanol was the superior cryopreservation method for Chlamydomonas compared to the commercial kit and that post-thaw culture conditions dramatically affect viability. We also demonstrated that cryopreserved cells could be successfully thawed and plated directly onto solid agar plates. Our findings have important implications for the long-term storage of Chlamydomonas that can likely be extended to other algal species.
authors.affiliation: University of Mississippi Medical Center, Department of Biochemistry, Guyton Building, G225, 2500 N. State St., Jackson, MS 39216, USA. Electronic address: cscarbrough@umc.edu.
authors.forename: Chasity
authors.initials: C
authors.lastname: Scarbrough
authors.affiliation: University of Mississippi Medical Center, Department of Biochemistry, Guyton Building, G225, 2500 N. State St., Jackson, MS 39216, USA. Electronic address: mwirschell@umc.edu.
authors.forename: Maureen
authors.initials: M
authors.lastname: Wirschell
mesh_terms: D016825:Chlamydomonas reinhardtii; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D003470:Culture Media
chemical_list: D003451:Cryoprotective Agents; D003470:Culture Media
keywords: Algae; Chlamydomonas; Cryopreservation; GeneArt; Methanol