I wondered if someone had ever stored spore suspension at -80C just in water or a saline solution without any glycerol or cryopreservation agent.
We would like to transfer a protocol to LMICs to prepare first-line controls for C. perfringes assays. Purchasing glycerol for stock cultures has been problematic in the past (the purchased solution was not of good quality, leading to spores death). A colleague has suggested going without the addition of any glycerol, but I can´t find literature with pros or cons.
Thanks for your insights!
Glycerol is widely used in microbial preservation because it prevents ice crystal formation, which can damage cellular structures during freezing. Without glycerol, spores may be more vulnerable to damage at low temperatures. Preserving Clostridia spores at -80°C in water without glycerol presents several challenges. Although Clostridia spores are known for their resilience, the lack of glycerol—a cryoprotectant—may affect their viability during freezing and thawing processes. While these spores can withstand various environmental stresses, including extreme temperatures, studies indicate that the absence of a cryoprotectant could lead to lower recovery rates post-thawing. If glycerol is not an option, consider alternative cryoprotectants such as dimethyl sulfoxide (DMSO) or trehalose, which may offer some protection without compromising spore integrity. Additionally, storing the spores in a buffered saline solution instead of plain water could help maintain their viability during freezing.
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