celltrace blue cell proliferation kit c34568  (Thermo Fisher)

Journal: Cell Reports Medicine

Article Title: Immunomodulatory effects and improved outcomes with cisplatin- versus carboplatin-based chemotherapy plus atezolizumab in urothelial cancer

doi: 10.1016/j.xcrm.2024.101393

Figure Lengend Snippet: Co-culture of cisplatin- versus carboplatin-pretreated MC38 cancer cells with bone marrow-derived DCs and OT-I T cells induces DC activation, OT-I T cell proliferation, and antigen-specific T cell-mediated MC38 killing (A) HMGB1 secretion as measured in the supernatant collected from MC38 cancer cells treated with increasing concentrations of cisplatin (red) or carboplatin (black). Supernatant was collected 3 days after treatment. Data depict one representative experiment of two independent experiments; duplicate conditions for each experiment. Data are mean ± SEM. (B) Protein expression of MHC-I, PD-L1, CD40, and CD86 as measured by median fluorescence intensity in monocyte-derived DCs after 24 h of co-culture with MC38 cancer cells that were primed with dose titrations of cisplatin (red) or carboplatin (black) for 24 h. Data depict one representative experiment of three independent experiments; duplicate conditions for each experiment. Data are mean ± SEM. (C) Flow cytometry histograms depicting OT-I T cell proliferation measured by CellTrace Blue dilution assay. MC38-OVA cells were pretreated with dose titrations of cisplatin or carboplatin for 24 h. OT-I T cells were co-cultured with pretreated MC38-OVA cells for 3 days in the presence or absence of monocyte-derived DCs before the proliferation assay. Data depict one representative experiment of two independent experiments. (D and E) Representative flow cytogram showing OT-I T cell-mediated MC38-OVA cancer cell killing (D) and the summarized percentage of OT-I T cell-mediated tumor cell death (E) as measured by 7-AAD and annexin V staining. MC38-OVA tumor cells were pretreated with increasing concentrations of cisplatin or carboplatin for 24 h, followed by co-culture with OT-I T cells for 5 h before the assay. Data depict one representative experiment of three independent experiments; duplicate conditions for each experiment. Data are mean ± SEM. See also Figure S9 .

Article Snippet: The following day, OT-I CD8 + T cells labeled with the CellTrace Blue Cell proliferation kit (Thermo Fisher Scientific, Cat# C34568) were added at a density of 50,000 cells/well in the presence or absence of DCs.

Techniques: Co-Culture Assay, Derivative Assay, Activation Assay, Expressing, Fluorescence, Flow Cytometry, Dilution Assay, Cell Culture, Proliferation Assay, Staining

Journal: Cell Reports Medicine

Article Title: Immunomodulatory effects and improved outcomes with cisplatin- versus carboplatin-based chemotherapy plus atezolizumab in urothelial cancer

doi: 10.1016/j.xcrm.2024.101393

Figure Lengend Snippet:

Article Snippet: The following day, OT-I CD8 + T cells labeled with the CellTrace Blue Cell proliferation kit (Thermo Fisher Scientific, Cat# C34568) were added at a density of 50,000 cells/well in the presence or absence of DCs.

Techniques: Blocking Assay, Recombinant, Cell Isolation, Activation Assay, Enzyme-linked Immunosorbent Assay, Staining, Software