DLBCL cells with ferroptosis morphology can be detected with a deep convolutional neural network.

It has been demonstrated that diffuse large B-cell lymphoma (DLBCL) is especially sensitive to ferroptosis. Currently, confirming the presence of ferroptosis requires flow cytometry, which is a time consuming and labor-intensive task. Blistering of the cell membrane has been shown to be a ferroptosis-specific morphological change. In this study we developed a deep convolutional neural network to detect the blistering of cell membrane. Buthionine sulfoximine treatment increased the percentage of blistering cells from 2 % to 38 % (p < 0.001) when glutathione was deprived from the culture media. Ferrostatin-1 treatment completely reversed the effect. Imidazole ketone erastin (IKE) and auranofin treatment increased blistering cells gradually in dose response manner from 5.4 % to 18.1 % (p < 0.05) and 6.1-50.1 % (p < 0.0001) respectively. We also tested malignant melanoma and breast cancer cell lines to confirm that the blistering phenomena can also be observed in adherent cell lines. We used fluorescence-activated cell sorting to measure the lipid peroxidation associated with ferroptosis and found a significant increase of bodiby-C11 _oxidized mean compared to DMSO controls for IKE (345 vs 462, p < 0.01) and auranofin (345 vs 686.5, p < 0.05).
Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Pyry Kotkaranta reports financial support was provided by Terttu Foundation. Mikko Chan reports financial support was provided by Terttu Foundation. Milla E.L Kuusisto reports financial support was provided by Terttu Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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