After collecting human feces, I want to perform mouse fecal microbiota transplantation. Can I directly freeze the collection tube at -80°C, or do I need to add glycerol or similar substances for freezing?
In mouse fecal microbiota transplantation (FMT) experiments, the freezing and handling of fecal samples are crucial for the survival of microbial communities and the subsequent transplantation effect. Directly freezing the sample collection tube at -80°C may lead to decreased viability and diversity of gut microbiota cells, so protective measures are usually taken to maintain microbial viability and integrity. The standard method for freezing fecal samples is as follows:
1. Cryoprotectants: To protect anaerobic bacteria and other microorganisms in the feces, cryoprotectants are usually added, such as:
(1) Glycerol: The final concentration is usually 10%-20%. Glycerol can effectively prevent ice crystal formation and reduce cell membrane damage.
(2) DMSO (Dimethyl Sulfoxide): Used at a concentration of 5%-10%. It also has protective effects but can be toxic to some bacteria, so it is used less frequently.
(3) Bovine Serum Albumin (BSA): Sometimes 1%-5% BSA is added to enhance the protective effect.
2. Procedure
(1) Suspend fresh fecal samples evenly in a cryoprotective solution (such as 10% glycerol) under anaerobic conditions.
(2) Dispense into sterile cryogenic vials under aseptic conditions, usually 200-500μL per vial.
(3) Quickly freeze at -80°C and avoid repeated freeze-thaw cycles.
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