ls174t atcc cl  (ATCC)

Journal: Theranostics

Article Title: Three-dimensional colon cancer organoids model the response to CEA-CD3 T-cell engagers.

doi: 10.7150/thno.63359

Figure Lengend Snippet: Figure 1. Modelling cytotoxicity of CD3-CEA bispecific T-cell engagers of different affinity on 3D tumor spheroids. (A) Representative flow cytometry histograms of CEA surface expression on LS174T cells. (B) Representative images of cocultures of LS174T spheroids (Red) with T cells (Blue) and the CEA-TCB, CEACAM5-TCB T-cell engagers and an irrelevant mouse IgG1 control (Ctrl). Active caspase 3 is shown by a fluorescent probe (Green) that marks apoptotic tumor cells (Bars 50 µm). (C) Quantification of apoptotic cells in images as in B and videos as Video S1. (D) Representative flow cytometry histogram of CEA surface expression on HT29 cells. (E) Representative images of cocultures of HT29 spheroids (Red) with T cells (Blue) and the CEA-TCB, CEACAM5-TCB T-cell engagers and an irrelevant mouse IgG control (Ctrl). Active caspase 3 is shown by a fluorescent probe (Green) that indicates apoptotic tumor cells (Bars 50, 20 µm). (F) Quantification of apoptotic cells in images as in E and videos as Video S1. Representative experiments are shown out of at least two rendering similar results. Quantifications in C and F were performed analyzing four to seven spheroids per condition. Means±SD are shown for quantitative data. * p<0.05; U Mann Whitney tests were performed for statistical comparisons.

Article Snippet: LS174T human colon carcinoma cells (ATCC CL 188), parental, modified to stably express red fluorescent protein (RFP) have been published elsewhere [12] and were cultured in DMEM (Gibco/Life Technologies) supplemented with 10% FCS, under standard cell culture conditions (37 °C, 5% CO2).

Techniques: Flow Cytometry, Expressing, Control, MANN-WHITNEY

Journal: Theranostics

Article Title: Three-dimensional colon cancer organoids model the response to CEA-CD3 T-cell engagers.

doi: 10.7150/thno.63359

Figure Lengend Snippet: Figure 5. Co-cultures of tumor organoids with autologous FAP+ fibroblasts do not impair CEA redirected killing by T cells. (A) Representative images of a coculture of tumor cell organoids (Red) and tumor-associated fibroblasts (Green) from the patient 47389 with allogeneic T cells. A 3D reconstruction and a single plane of the same two organoids are shown to better visualize the interactions of tumor cells with fibroblasts. (B) Series of Z-stack planes of an organoid/fibroblast/T-cell co-culture as in A to show close contact of fibroblasts with organoids. (C) Time-lapse sequence of a triple Organoid/fibroblast/T-cell coculture treated with CEA-TCB antibodies and showing tumor cell cytotoxicity by activated caspase 3. (D) LS174T spheroids were set up in cocultures with T cells and CEA-TCB antibody or IgG1 irrelevant control antibody (Ctrl) and different amounts of fibroblasts (0,5x103, 1.5x104, 3x104 fibroblasts) were added to the culture. Numbers of recovered tumor cells assessed by flow cytometry after 24 hours of coculture are shown. (E) Representative flow cytometry histogram of fibroblast activated protein (FAP) expression on fibroblast in cultures from patients 47389 and 47550. (F) CD137 induction on CD8 T cells in cocultures as in D as measured by flow cytometry.

Article Snippet: LS174T human colon carcinoma cells (ATCC CL 188), parental, modified to stably express red fluorescent protein (RFP) have been published elsewhere [12] and were cultured in DMEM (Gibco/Life Technologies) supplemented with 10% FCS, under standard cell culture conditions (37 °C, 5% CO2).

Techniques: Co-Culture Assay, Sequencing, Control, Flow Cytometry, Expressing