Just wondering if there is a maximum time limit before lymphocyte viability is severely hindered after mouse death. For example, sometimes an immunization can result in a negative outcome (i.e. mouse death) before downstream work is scheduled to begin. If this happens it's best to harvest lymph nodes as soon as possible and proceed with the required downstream work. I'm just not sure what that maximum time frame would be post-death that would still result in viable lymphocytes being isolated. Thanks.
Hello Joanne Smith
Lymphocytes will begin to degrade upon mouse death. The sooner the isolation, the better the chances of maintaining cell viability and function. While some studies suggest that lymphocyte viability can be maintained for a few hours after death if kept on ice, aiming for isolation within 30 minutes to an hour after sacrifice is generally recommended for optimal results.
One study (an autopsy-based study) has revealed that morphology of all WBCs was found to be normal till 6 hours until death (paper attached below).
https://ijop.net/index.php/mlu/article/view/762
Best,
Joanne Smith , lymph nodes should be collected and single-cell suspensions prepared as soon as possible after death (ideally within 30 minutes, no more than three hours, and kept cold throughout). Otherwise, the viability of most lymphocytes will be severely compromised, making them unsuitable for reliable experimental use.
- Two publications provide clear instructions for lymph node sampling: PMID: 29553537 PMID: 27842342
- The Protocol below:
- https://escholarship.org/content/qt0w27n5zt/qt0w27n5zt_noSplash_9297f6a8ac0bdf25ea69fd6a20eed8f4.pdf
The answer to this question comes from MedChemExpress (MCE) Technical Support.
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