Hello,
I am interested in isolating TILs from bladder cancer murine tumors injected into C57BL/6 mice. I wonder what is the best way to do it in order to have a good yield of cells (just interested in running flow to know numbers of TILs and will not be doing any assays using the cells isolated)
-Should I use Enzymatic digestion or it will affect cells isolated?
-Is it better to use ficoll gradient to get rid of tumor debris?
Your answers will be greatly appreciated
You can consider manual microdissection of the TIL area under dissection microscope.
We always use the GentleMacs Dissociator together with enzymatic digestion and have consistent and replicable data with B16 melanomas (s.c. injection). Using Ficoll can also give you good results. However, in my experience especially if there is a lot of debris then I find a lot of living cells in the pellet or a lot of debris still in the intermediate fraction.
For the assessment of TIL numbers, we use the built-in FlowSensor of our BD FACSVerse but the addition of beads should also work well.
I agree with Wijendra Senarathne. For highest precision, I would recommend a laser microdissection system (such as MMI's CellCut), to be able to selectively isolate single cells and cell cluster.
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