Can someone recommend a good solution and tube material to freeze aav2/9 in a -80C freezer? Is a cryoprotectant required?
We currently froze our AAV product in DPBS(+Cacl2,+MgCl2) into a cryovial but I've been reading articles that recommend all sorts of freezing methods.
hi
Freezing and storage of adeno-associated virus (AAV) can affect the stability and viability of the virus, and therefore it is important to carefully consider the method and conditions used for freezing and storage.
One recommended method for freezing AAV is to use a cryoprotectant solution, such as dimethyl sulfoxide (DMSO) or glycerol, to protect the virus from damage during the freezing process. AAV can be sensitive to osmotic stress, and the presence of a cryoprotectant can help to prevent the formation of ice crystals and to maintain the structural integrity of the virus.
To prepare an AAV solution for freezing, it is generally recommended to use a buffer such as phosphate-buffered saline (PBS) or Dulbecco's modified Eagle's medium (DMEM) to solubilize the virus. The buffer should be supplemented with calcium and magnesium ions to support the stability of the virus, and the pH should be adjusted to a neutral or slightly basic range. The AAV solution can then be mixed with a cryoprotectant, such as DMSO or glycerol, at a final concentration of 10-30% (v/v). The solution can then be aliquoted into tubes or vials, sealed tightly, and stored in a -80°C freezer.
It is important to note that the choice of cryoprotectant, concentration, and storage temperature may affect the stability and viability of AAV, and it is recommended to optimize these conditions for each specific AAV serotype and application. The use of a cryoprotectant is generally recommended to prevent damage to the virus during the freezing process, but it may not be necessary in all cases. For example, some AAV serotypes may be more stable and resistant to freezing damage than others, and may not require the use of a cryoprotectant.
In terms of the material of the tubes or vials used for freezing, it is generally recommended to use tubes or vials made of materials that are resistant to breakage and do not interact with the AAV solution. Glass or plastic tubes or vials can be used, but it is important to carefully consider the compatibility of the material with the AAV solution and the cryoprotectant. Glass tubes or vials may be more resistant to breakage, but they may be more prone to contamination and may interact with certain cryoprotectants. Plastic tubes or vials may be more prone to breakage, but they are generally more resistant to contamination and are compatible with a wider range of cryoprotectants. It is recommended to choose a tube or vial material that is suitable for the specific requirements of your study and to properly seal and label the tubes or vials to prevent contamination and ensure proper storage.
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