Can you store lentivirus in liquid nitrogen?

Dear Esteemed Colleague,

Greetings. I trust this message finds you well and advancing in your valuable research endeavors, particularly in the domain of lentiviral vector studies. Your inquiry regarding the storage of lentivirus in liquid nitrogen is both relevant and crucial for maintaining the integrity and efficacy of lentiviral preparations. Below, I provide a comprehensive analysis of lentivirus storage practices, with a focus on the use of liquid nitrogen.

Storage of Lentivirus

Lentiviruses are versatile tools in molecular biology and gene therapy research, used for delivering genetic material into cells. Preserving their infectious and functional capacities through proper storage is essential for the success of experimental outcomes.

Liquid Nitrogen Storage

Feasibility: Yes, lentiviruses can be stored in liquid nitrogen. Storing lentivirus at the temperature of liquid nitrogen (-196°C) is a method used to preserve the viral particles for long-term storage, minimizing the risk of degradation and loss of infectivity.

Preparation for Storage:Prior to storage, lentiviral stocks should be aliquoted in cryo-safe vials to avoid repeated freeze-thaw cycles that can significantly reduce viral titer and infectivity. It is advisable to mix the lentivirus aliquots with a cryoprotectant, such as sterile glycerol or DMSO, to a final concentration of 5-10%. This helps protect the viral particles from damage during the freezing process.

Process:Carefully label each vial with relevant information, including the date of preparation, viral titer, and any genetic modifications. Place the aliquoted vials in a controlled-rate freezing container or use a styrofoam box to achieve a gradual temperature decrease before transferring them to the liquid nitrogen storage tank.

Considerations and Best Practices

Safety Precautions: Given the biohazardous nature of lentiviruses, ensure that all procedures for handling, aliquoting, and storage comply with institutional biosafety regulations. Wear appropriate personal protective equipment and use biological safety cabinets.
Inventory Management: Maintain a detailed inventory of the stored lentiviral aliquots, including their specific locations within the liquid nitrogen storage system, to facilitate easy retrieval and avoid cross-contamination.
Thawing for Use: When retrieving lentivirus from liquid nitrogen storage, quickly thaw the aliquot in a 37°C water bath and immediately proceed to the intended application to maximize viral integrity and infectivity.
Alternative Storage Options: While liquid nitrogen offers an option for long-term storage, lentiviruses can also be stored at -80°C for several months without significant loss of infectivity, provided that freeze-thaw cycles are minimized.

Conclusion

Storing lentivirus in liquid nitrogen represents a viable strategy for preserving viral integrity and infectivity over extended periods. Adhering to meticulous preparation, safety guidelines, and proper cryopreservation techniques will ensure the lentivirus remains a potent tool in your research endeavors.

Should you require further insights into lentiviral storage practices, handling, or applications, please do not hesitate to reach out. I am here to support your research efforts and facilitate your success in the field of lentiviral vector studies.

Warm regards.

This list of protocols might help us better address the issue.

Arvind Singh

Please check the following ResearchGate link.

https://www.researchgate.net/post/Has_anyone_concentrated_and_frozen_aliquoted_lentivirus

Shirley Wu

Thanks, Arvind! I am more interested in whether lentivirus can be stored in liquid nitrogen in the long term. The link you posted was mainly about freeze thaw cycles and storage at -80. They did mention snap freeze in liquid nitrogen but not long term storage. Thanks anyway!

Wajid Nasim Jatoi

Dear Shirley Wu,

If you are planing to use the virus for multiple experiments, aliquot the volume that is required, and in the form of liquid nitrogen/a dry ice/ethanol bath before high @ -80°C. Lentivirus existance at -80°C varies depending on the media composition and the virus determined.

Thanks!

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