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CTLL-2 is a mouse T-cell lymphoma cell line that can be used in various cell culture and research applications. The cell line is derived from a T-cell lymphoma induced in a C57BL/6 mouse. CTLL-2 cells are interleukin-2 (IL-2) dependent and can be used to study T-cell activation and proliferation.

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2 protocols using ctll 2

1

MUC1-Expressing Tumor Cell Lines for Preclinical Studies

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All cell culture reagents were purchased from Biowest and Serana. Tumor cell lines ZR-75-1, MCF-7, HSC-4, CaoV-3 and T-47D as well as the human NK cell line KHYG-1 and murine T cell line CTLL-2 were obtained from DSMZ or ATCC and cultured in the respective recommended medium under standard conditions. KHYG-1 and CTLL-2 were cultured in presence of 10 ng/mL IL-2 (PeproTech, Rocky Hill, NJ, USA).
For in vivo studies, the murine tumor cell lines B16.F10 and CT26.wt (ATCC) were stably transfected by electroporation (Amaxa) with a plasmid coding for human MUC1 (40 tandem repeats) and confirmed for TA-MUC1 expression in vitro and in vivo in mice by flow cytometry and immunohistochemistry using GT-00A. Cells were cultured in standard medium +100 nM methotrexate (Hexal, Holzkirchen, Germany) as selection pressure. For the metastasis model, hMUC1-B16.F10 cells were further transduced to stably express firefly luciferase (110 RLU/cell) enabling detection of metastasis. Origin, TA-MUC1 and IL-15R expression status of cell lines are indicated in Table S1.
Peripheral blood mononuclear cells (PBMCs) were prepared from commercially available buffy coats (DRK Berlin) or leukapheresates (Charité Berlin) of healthy donors by Biocoll separation (Biochrom, Cambridge, UK) and density gradient centrifugation. PBMCs from leukapheresate products were stored frozen in liquid nitrogen.
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CTLL-2 Cell Proliferation Assay

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CTLL-2 cells (ATCC) were cultured according to ATCC's protocol. For the CTLL-2 proliferation assay, cells were grown to a density of 1 × 105cells/mL. Duraleukins or hIL-2 (PeproTech) was added into the wells of a flat-bottom 96-well tissue culture microplate. The final concentration of hIL-2 and Duraleukins was adjusted to 0.01–25 ng/mL and 0.025–50 ng/mL by 2-fold serial dilution. Every concentration group was tested in duplicate. 104 cells were seeded to each well of the plate, and the total volume in each well was 100 μL. The cells were then incubated at 37°C, 5% CO2 for 56 h. 10 μL of WST-1 reagent (Roche) was added to each well and the cells were incubated for another four h. The plate was shaken at 1,000 rpm for 1 min before analysis. A spectrometer read absorbance at 450 nm. 690 nm was used as a reference wavelength. A dose-response curve was fit via nonlinear regression using GraphPad Prism 7.
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