Generating Mouse Organoids and opT Cells

Mouse organoids were generated from triple-negative breast cancer mouse models.21 (link) Autologous mouse T cells from the spleen were isolated by EasySep Mouse T Cell Isolation Kit (Stemcell Technologies, 19851). To generate opT, mouse organoids, and T cells were cultured in mouse T cell medium: Serum-free medium (CellGernix, 20 801–0100), 10% fetal bovine serum (FBS) (Glico, 16140071), mouse interleukin (IL)-2 (1000 IU/mL) (Peprotech, 400–02), mouse IL-15 (10 ng/mL, Peprotech, 400–24) and mouse IL-21 (10 ng/mL, Prospec, CYT-033). For killing assay, the organoids from mouse tumors or normal tissue were co-cultured with opT cells. The supernatants from the co-culture were harvested, and secreted IFN-γ was quantified by IFN-γ ELISA kit (Mabtech, 3421–1 H-6). Please see online supplemental methods for further details.

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Meng Q., Xie S., Gray G.K., Dezfulian M.H., Gandarilla O., Li W., Huang L., Akshinthala D., Ferrer E., Conahan C., Perea Del Pino S., Grossman J., Elledge S.J., Hidalgo M, & Muthuswamy S.K. (2021). Empirical identification and validation of tumor-targeting T cell receptors from circulation using autologous pancreatic tumor organoids. Journal for Immunotherapy of Cancer, 9(11), e003213.

Publication 2021

EasySep Mouse T Cell Isolation Kit mouse IL-15 IFN-γ ELISA kit

Assay Cell isolation Co culture Cultured cells Elisa Fetal bovine serum Ifn γ Interleukin 15 Interleukin 2 Interleukin 21 Isolation Mouse Normal tissue Organoids Prospec Serum Spleen Stemcell T cell Triple negative breast cancer Tumors

Corresponding Organization : Cornell University

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Variable analysis

independent variables

Mouse organoids generated from triple-negative breast cancer mouse models
Autologous mouse T cells from the spleen

dependent variables

Secreted IFN-γ quantified by IFN-γ ELISA kit

control variables

Mouse T cell medium: Serum-free medium, 10% fetal bovine serum (FBS), mouse interleukin (IL)-2 (1000 IU/mL), mouse IL-15 (10 ng/mL), and mouse IL-21 (10 ng/mL)

controls

Positive control: Co-culture of organoids from mouse tumors with opT cells
Negative control: Co-culture of organoids from normal tissue with opT cells

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