Skip to main content
. 2018 Dec 3;110(1):269–278. doi: 10.1111/cas.13873

Figure 2.

Figure 2

Cancer‐associated fibroblasts (CAF) and conditioned media support survival of primary lymphoma cells. A, Scheme of coculture of primary lymphoma cells with CAF. B, Proliferation of primary lymphoma cells (Pt#8, left; Pt#9, right) in monoculture (open circle) and coculture with CAF#1 (circle) and CAF#2 (square). Survival of tumor cells was evaluated by Trypan blue exclusion. Each point represents the mean value taken from three independent experiments with error bars indicating the standard error of the mean. C, Immunoblotting for γH2A.X, CC3, and GAPDH as a loading control in Pt#8 and Pt#9 in monoculture and coculture with CAF#1 and CAF#2 was carried out. D, Viability of three histological types of primary B‐cell lymphoma cells (DLBCL, indigo blue; FL, green; MZBCL, brown) in coculture with CAF#1 (left) and CAF#2 (right) overnight are shown. Each point represents the mean value of two technical replicates taken from a single experiment. E, Viability of a different lineage of lymph node samples including B‐cell lymphoma (indigo blue), T‐cell lymphoma (green) and others such as related disease or reactive tissue (brown) in coculture with CAF#1 and CAF#2 overnight are shown. Each point represents the mean value of two technical replicates taken from a single experiment. F, Scheme of culture in the presence of conditioned medium (CM) released from CAF (left), in the absence of direct contact of primary lymphoma cells to CAF by Transwell (upper right), and coculture with CAF (lower right). G, Comparison of survival of primary B‐cell lymphoma cells (Pt#2, Pt#5, Pt#1, and Pt#6) in monoculture (white), coculture with CAF (orange), in the absence of direct contact with CAF using the Transwell (green), and in culture with CM released from CAF (blue) overnight are shown. CAF originated from samples corresponding to the examined lymphoma cells. Each bar represents averaged data (two technical replicates) taken from a single experiment. H, Comparisons of redox regulation in B‐cell lymphoma cells (Pt#2, Pt#5, Pt#1, and Pt#6) in monoculture (white), coculture with CAF (orange), in the absence of direct contact with CAF using Transwell (green), and in culture with CM released from CAF (blue) are shown. Levels of intracellular ROS were measured using a fluorescent indicator, 2′,7′‐dichlorofluorescin diacetate. I, Comparisons of redox regulation in B‐cell lymphoma cells (Pt#2, Pt#5, Pt#1, and Pt#6) in monoculture, and cultures with CM released from CAF#2, CAF#5, CAF#1, and CAF#6. Blue bars represent mean values for the four conditions. Values of the matched pairs of lymphoma cells and established CAF are indicated as black points, and those of unmatched pairs are indicated as grey points. *P < .05, **P < .01, ***P < .001. J, Survival of primary B‐cell lymphoma cells (Pt#2, Pt#5, Pt#1, and Pt#6) in culture with CM released from the corresponding CAF. Blue bar represents the mean value of the four conditions (Pt#2, Pt#5, Pt#1, and Pt#6) indicated by black points. K, Survival of primary lymphoma cells (Pt#9) in the presence of 100 nmol/L doxorubicin in culture with or without CM released from CAF#2. Cells were treated with doxorubicin for 72 h, before analysis of cell death on flow cytometry. Each bar represents averaged data (two technical replicates) taken from five independent experiments. Error bars indicate SD