Hello everyone, I am trying to plan logistics and experiments using human tissues. The study is multicenter based and I will be receiving samples from different centers. The transport time should be around 48h. My plan is to digest the tissues on arrival and run flow for different immune cell populations. Has anyone experience with such transport and cell viability? Is there any special transport media or RPMI is enough? Transport temperature? Looking forward to suggestions. Thanks !!
Hello Viqar,
This is highly dependent on the tissue type and the magnitude of immune cell infiltration in it. Generally, RMPI is enough and keeping the tissue on ice during transportation is important for cell viability. You can ask the supplying center to dice the tissue into small pieces to ensure better exposure to media. I used to receive tissue at the very end of the day, dice them and keep them at 4C to the next day to process and that was OK for breast cancer tissue. However, I did confirm that this was OK for what I was looking for by splitting the same tissue and processing one half immediately and the other the next day before I move on with optimized protocol.
The most important thing is to be consistent in treating your tissues to the best you can and I would suggest trying different protocols till you set an optimized one that works best for you.
Best,
Cells/tissues need serum to keep them happy; so using just RPMI may not be a good idea esp. when you have 48 hours lag time. However, since you are working with human samples which I assume are precious, I would recommend that you use serum free media like AIM-V or X-VIVO 10 esp. when you plan to look at immune cell subsets. To add to what Kawther K. Ahmed said, Miltenyi seems to have a storage solution that they claim could prevent unwanted changes in cell signaling pathways during storage/shipment. You could maybe try that out in addition to media and select the conditions that work the best in your experimental setup.
Hi,
I suggest you conduct a pilot simulation experiment in your lab; get the tissue of interest, treat and keep in your lab at the expected storage conditions over the anticipated period of transportation and then run your flow and viability on ViCell, for instance. This should give you an idea of what to expect, in the event you couldn't get a perfect solution a priori.
Best,
Kawther K. Ahmed and Aminu Abba Yusuf Thank you for your suggestions. I would run a small set of pilot experiments. I am in the process of setting up a protocol based on previous experiences in the lab.
Julie Joseph Thanks for suggesting serum free media. You just answered my follow up question! would be interesting to compare the newer serum free media and the classic RPMI+5% FBS as a colleague suggested. Cheers!!
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