I would appreciate any helpful information regarding CFSE labeling for in-vivo vs. in-vitro experiments. Below my client describes his results: In the in vivo experiments, I inject (intravenously) CFSE-labeled CD4+ T cells from C57B/6 mouse into bm12 mouse (allogenic setting). When I pull out the splenocytes and lymph nodes from the bm12 mouse and run flow analysis, the CFSE proliferation was not observed 48 h post-stimulation, but from 72 h onwards, CFSE proliferation was consistant (7 rounds of proliferation; undivided cells about 30%; precursor frequency about 14%). When I tried to repeat the same experiment in vitro, I see a prominant bunch of cells that are low on CFSE scale (corresponding to 4-5 rounds of division) in addition to the undivided (CFSE high) peak within 48 h. I do not see any ordered CFSE proliferation in vitro further on (after 48 h). My CFSE + mimosin and CFSE + ConA controls are very fine. We wonder whether synchronious stimulation in vitro is the reason for this observation, or whether I am missing something somewhere. Thanks in advance! Paul L. Hallberg Flow Cytometry Manager KCC CORE Flow Cytometry Facility Thomas Jefferson University 215-503-4556 215-923-0249(fax) Paul.Hallberg@mail.tju.edu
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