Establishment of imageable model of T-cell lymphoma growing in syngenic mice.
BACKGROUND
Cancer research is focused on processes which influence in vivo tumor growth dynamics, tumor microenvironment and antitumor immune responses. Recently, it was documented that some cytostatics, including their polymeric derivatives, were able to trigger an anticancer immune response. Such interactions are studied mainly in vitro but relevant in vivo studies are necessary and were only recently started. Whole-body imaging of fluorescently labeled tumors, which enables visualization of the whole-body down to single cell-cell interactions, is therefore a promising tool in understanding these processes.


METHODS
EL-4 T-cell lymphoma cells were transfected with plasmid containing either fusion construct of enhanced green fluorecent protein (EGFP) with H2B histone or pure EGFP gene under cytomegalovirus promotor and resistance to neomycin. Stability of expression was determined by flow cytometry and cellular localization of green fluorescence signal was tested using fluorescent microscopy. An in vivo whole-body imaging system was used to evaluate growth in vivo.


RESULTS
EL-4 cells were successfully transfected and established stable transfectants with a proliferation rate comparable to that of wild-type EL-4. Clone 12, with very strong whole-cell expression, enables tracking of metastatic spreading, whereas clone 3, with EGFP within the cell nucleus, allows frozen section analysis and observing of interaction with immunocompetent cells.


CONCLUSIONS
Established imageable EL-4-EGFP(+) cell lines are a magnificent tool for the study of tumor growth and the tumor microenvironment.
Vetvicka
David
D
Institute of Microbiology AS CR, Videnska 1083, 142 00 Prague 4, Czech Republic. vetvicka@biomed.cas.cz
Hovorka
Ondrej
O
Kovar
Lubomir
L
Rihova
Blanka
B
D000818:Animals; D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D005456:Fluorescent Dyes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006657:Histones / Q000235:genetics; D016399:Lymphoma, T-Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D014162:Transfection
D005456:Fluorescent Dyes; D006657:Histones; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins